TRANSPOSON SYSTEM AND METHODS OF USE

INCORPORATION OF SEQUENCE LISTING

The contents of the text file named “POTH-029/001WO_SeqList.txt,” which was created on Aug. 31, 2018 and is 44,366 KB in size, are hereby incorporated by reference in their entirety.

FIELD OF THE DISCLOSURE

The present invention is directed to compositions and methods for targeted gene modification.

BACKGROUND

Ex vivo genetic modification of non-transformed primary human T lymphocytes using non-viral vector-based gene transfer delivery systems has been extremely difficult. As a result, most groups have generally used viral vector-based transduction such as retrovirus, including lentivirus. A number of non-viral methods have been tested and include antibody-targeted liposomes, nanoparticles, aptamer siRNA chimeras, electroporation, nucleofection, lipofection, and peptide transduction. Overall, these approaches have resulted in poor transfection efficiency, direct cell toxicity, or a lack of experimental throughput.

The use of plasmid vectors for genetic modification of human lymphocytes has been limited by low efficiency using currently available plasmid transfection systems and by the toxicity that many plasmid transfection reagents have on these cells. There is a long-felt and unmet need for a method of nonviral gene modification in immune cells.

SUMMARY

When compared with viral transduction of immune cells, such as T lymphocytes, delivery of transgenes via DNA transposons, such as piggyBac and Sleeping Beauty, offers significant advantages in ease of use, ability to delivery much larger cargo, speed to clinic and cost of production. The piggyBac DNA transposon, in particular, offers additional advantages in giving long-term, high-level and stable expression of transgenes, and in being significantly less mutagenic than a retrovirus, being non-oncogenic and being fully reversible. Previous attempts to use DNA transposons to deliver transgenes to T cells have been unsuccessful at generating commercially viable products or manufacturing methods because the previous methods have been inefficient. For example, the poor efficiency demonstrated by previous methods of using DNA transposons to deliver transgenes to T cells has resulted in the need for prolonged expansion ex vivo. Previous unsuccessful attempts by others to solve this problem have all focused on increasing the amount of DNA transposon delivered to the immune cell, which has been a strategy that worked well for non-immune cells. This disclosure demonstrates that increasing the amount of DNA transposon makes the efficiency problem worse in immune cells by increasing DNA-mediated toxicity. To solve this problem, counterintuitively, the methods of the disclosure decrease the amount of DNA delivered to the immune cell. Using the methods of the disclosure, the data provided herein demonstrate not only that decreasing the amount of DNA transposon introduced into the cell increased viability but also that this method increased the percentage of cells that harbored a transposition event, resulting in a viable commercial process and a viable commercial product. Thus, the methods of the disclosure demonstrate success where others have failed.

The disclosure provides a nonviral method for the ex-vivo genetic modification of an immune cell or an immune cell precursor comprising delivering to the immune cell or the immune cell precursor, (a) a nucleic acid or amino acid sequence comprising a sequence encoding a transposase enzyme and (b) a recombinant and non-naturally occurring DNA sequence comprising a DNA sequence encoding a transposon. In certain embodiments, the method further comprises the step of stimulating the immune cell or the immune cell precursor with one or more cytokine(s).

In certain embodiments of the methods of the disclosure, the sequence encoding a transposase enzyme is an mRNA sequence. The mRNA sequence encoding a transposase enzyme may be produced in vitro.

In certain embodiments of the methods of the disclosure, the sequence encoding a transposase enzyme is a DNA sequence. The DNA sequence encoding a transposase enzyme may be produced in vitro. The DNA sequence may be a cDNA sequence.

In certain embodiments of the methods of the disclosure, the sequence encoding a transposase enzyme is an amino acid sequence. The amino acid sequence encoding a transposase enzyme may be produced in vitro. A protein Super piggybac transposase (SPB) may be delivered following pre-incubation with transposon DNA.

In certain embodiments of the methods of the disclosure, the delivering step comprises electroporation or nucleofection of the immune cell or the immune cell precursor.

In certain embodiments of the methods of the disclosure, the method further comprises the step of stimulating the immune cell or the immune cell precursor with one or more cytokines. In certain embodiments, the step of stimulating the immune cell or the immune cell precursor with one or more cytokine(s) occurs following the delivering step. Alternatively, or in addition, in certain embodiments, the step of stimulating the immune cell or the immune cell precursor with one or more cytokine(s) occurs prior to the delivering step. In certain embodiments, the one or more cytokine(s) comprise(s) IL-2, IL-21, IL-7 and/or IL-15.

In certain embodiments of the methods of the disclosure, the immune cell or the immune cell precursor is an autologous immune cell or immune cell precursor. The immune cell or immune cell precursor may be a human immune cell, a human immune cell precursor, an autologous immune cell, and/or an autologous immune cell precursor. The immune cell may be derived from a non-autologous source, including, but not limited to a primary cell, a cultured cell or cell line, an embryonic or adult stem cell, an induced pluripotent stem cell or a transdifferentiated cell. The immune cell may have been previously genetically modified or derived from a cell or cell line that has been genetically modified. The immune cell may be modified or may be derived from a cell or cell line that has been modified to suppress one or more apoptotic pathways. The immune cell may be modified or may be derived from a cell or cell line that has been modified to be “universally” allogenic by a majority of recipients in the context, for example, of a therapy involving an adoptive cell transfer.

In certain embodiments of the methods of the disclosure, the immune cell is an activated immune cell.

In certain embodiments of the methods of the disclosure, the immune cell is a resting immune cell.

In certain embodiments of the methods of the disclosure, the immune cell is a T-lymphocyte. In certain embodiments, the T-lymphocyte is an activated T-lymphocyte. In certain embodiments, the T-lymphocyte is a resting T-lymphocyte.

In certain embodiments of the methods of the disclosure, the immune cell is a Natural Killer (NK) cell.

In certain embodiments of the methods of the disclosure, the immune cell is a Cytokine-induced Killer (CIK) cell.

In certain embodiments of the methods of the disclosure, the immune cell is a Natural Killer T (NKT) cell.

In certain embodiments of the methods of the disclosure, the immune cell is isolated or derived from a human.

In certain embodiments of the methods of the disclosure, the immune cell precursor is a stem cell or stem-like cell capable of differentiation into an immune cell. In some embodiments, the immune cell precursor is a hematopoietic stem cell (HSC). In some embodiments, the immune cell precursor is a primitive hematopoietic stem cell. In some embodiments, the immune cell precursor is a human HSC or human primitive HSC.

In certain embodiments of the methods of the disclosure, the method further comprising the step of differentiating the immune cell precursor into an immune cell. In some embodiments, the immune cell is a T lymphocyte (T cell), a B lymphocyte (B cell), a Natural Killer (NK) cell, or a Cytokine-induced Killer (CIK) cell.

In certain embodiments of the methods of the disclosure, the immune cell is isolated or derived from a non-human mammal. In certain embodiments, the non-human mammal is a rodent, a rabbit, a cat, a dog, a pig, a horse, a cow, or a camel. In certain embodiments, the immune cell is isolated or derived from a non-human primate.

In certain embodiments of the methods of the disclosure, the mRNA sequence encoding the transposase enzyme is produced in vitro.

In certain embodiments, the transposon is a piggyBac transposon or a piggyBac-like transposon. In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac transposon, the transposase is a piggyBac transposase. In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac-like transposon, the transposase is a piggyBac-like transposase.

In certain embodiments, the piggyBac transposase comprises an amino acid sequence comprising SEQ ID NO: 14487. In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac transposon, the transposase is a piggyBac™ or a Super piggyBac™ (SPB) transposase. In certain embodiments, and, in particular, those embodiments wherein the transposase is a Super piggyBac™ (SPB) transposase, the sequence encoding the transposase is an mRNA sequence.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac™ (PB) transposase enzyme. The piggyBac (PB) transposase enzyme may comprise or consist of an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14487)

1
MGSSLDDEHI LSALLQSDDE LVGEDSDSEI SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61
SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121
PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTGATFRD TNEDEIYAFF

181
GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV

241
FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RMYIPNKPSK YGIKILMMCD

301
SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ

361
EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC

421
DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481
SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPNEV

541
PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at one or more of positions 30, 165, 282, or 538 of the sequence:

In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at two or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at three or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at each of the following positions 30, 165, 282, and 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the amino acid substitution at position 30 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 165 of the sequence of SEQ ID NO: 14487 is a substitution of a serine (S) for a glycine (G). In certain embodiments, the amino acid substitution at position 282 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 538 of the sequence of SEQ ID NO: 14487 is a substitution of a lysine (K) for an asparagine (N).

In certain embodiments of the methods of the disclosure, the transposase enzyme is a Super piggyBac™ (SPB) transposase enzyme. In certain embodiments, the Super piggyBac™ (SPB) transposase enzymes of the disclosure may comprise or consist of the amino acid sequence of the sequence of SEQ ID NO: 14487 wherein the amino acid substitution at position 30 is a substitution of a valine (V) for an isoleucine (I), the amino acid substitution at position 165 is a substitution of a serine (S) for a glycine (G), the amino acid substitution at position 282 is a substitution of a valine (V) for a methionine (M), and the amino acid substitution at position 538 is a substitution of a lysine (K) for an asparagine (N). In certain embodiments, the Super piggyBac™ (SPB) transposase enzyme may comprise or consist of an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14484)

1
MGSSLDDEHI LSALLQSDDE LVGEDSDSEV SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61
SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121
PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTSATFRD TNEDEIYAFF

181
GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV

241
FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RVYIPNKPSK YGIKILMMCD

301
SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ

361
EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC

421
DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481
SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPKEV

541
PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF.

In certain embodiments of the methods of the disclosure, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 3, 46, 82, 103, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 258, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 486, 503, 552, 570 and 591 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 46, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 485, 503, 552 and 570. In certain embodiments, the amino acid substitution at position 3 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an asparagine (N) for a serine (S). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an alanine (A). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 82 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 103 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a serine (S). In certain embodiments, the amino acid substitution at position 119 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for an arginine (R). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) a cysteine (C). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 185 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 187 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for an alanine (A). In certain embodiments, the amino acid substitution at position 200 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 207 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a valine (V). In certain embodiments, the amino acid substitution at position 209 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a valine (V). In certain embodiments, the amino acid substitution at position 226 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a methionine (M). In certain embodiments, the amino acid substitution at position 235 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a leucine (L). In certain embodiments, the amino acid substitution at position 240 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 241 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 243 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a proline (P). In certain embodiments, the amino acid substitution at position 258 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an asparagine (N). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a proline (P). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine for a proline (P). In certain embodiments, the amino acid substitution at position 315 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for an arginine (R). In certain embodiments, the amino acid substitution at position 319 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a threonine (T). In certain embodiments, the amino acid substitution at position 327 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 328 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a cysteine (C). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 421 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for the aspartic acid (D). In certain embodiments, the amino acid substitution at position 436 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a valine (V). In certain embodiments, the amino acid substitution at position 456 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a methionine (M). In certain embodiments, the amino acid substitution at position 470 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 485 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a serine (S). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a methionine (M). In certain embodiments, the amino acid substitution at position 552 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 570 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a glutamine (Q). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a glutamine (Q).

In certain embodiments of the methods of the disclosure, the transposase enzyme is a Super piggyBac™ (SPB) transposase enzyme. The Super piggyBac (PB) transposase enzyme may comprise or consist of an amino acid sequence at least 75% identical to:

(SEQ ID NO: 14484)

MGSSLDDEHILSALLQSDDELVGEDSDSEVSDHVSEDDVQSDTEEAFI

DEVHEVQPTSSGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKH

CWSTSKSTRRSRVSALNIVRSQRGPTRMCRNIYDPLLCFKLFFTDEII

SEIVKWTNAEISLKRRESMTSATFRDTNEDEIYAFFGILVMTAVRKDN

HMSTDDLFDRSLSMVYVSVMSRDRFDFLIRCLRMDDKSIRPTLRENDV

FTPVRKIWDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFRVYIPNKP

SKYGIKILMMCDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPV

HGSCRNITCDNWFTSIPLAKNLLQEPYKLTIVGTVRSNKREIPEVLKN

SRSRPVGTSMFCFDGPLTLVSYKPKPAKMVYLLSSCDEDASINESTGK

PQMVMYYNQTKGGVDTLDQMCSVMTCSRKTNRWPMALLYGMINIACIN

SFIIYSHNVSSKGEKVQSRKKFMRNLYMSLTSSFMRKRLEAPTLKRYL

RDNISNILPKEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKANASCKK

CKKVICREHNIDMCQSCF.

In certain embodiments of the methods of the disclosure, the transposon is a Sleeping Beauty transposon. In certain embodiments of the methods of the disclosure, the transposase enzyme is a Sleeping Beauty transposase enzyme (see, for example, U.S. Pat. No. 9,228,180, the contents of which are incorporated herein in their entirety). In certain embodiments, the Sleeping Beauty transposase is a hyperactive Sleeping Beauty (SB100X) transposase. In certain embodiments, the Sleeping Beauty transposase enzyme comprises an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14485)

1
MGKSKEISQD LRKKIVDLHK SGSSLGAISK RLKVPRSSVQ TIVRKYKHHG TTQPSYRSGR

61
RRVLSPRDER TLVRKVQINP RTTAKDLVKM LEETGTKVSI STVKRVLYRH NLKGRSARKK

121
PLLQNRHKKA RLRFATAHGD KDRTFWRNVL WSDETKIELF GHNDHRYVWR KKGEACKPKN

181
TIPTVKHGGG SIMLWGCFAA GGTGALHKID GIMRKENYVD ILKQHLKTSV RKLKLGRKWV

241
FQMDNDPKHT SKVVAKWLKD NKVKVLEWPS QSPDLNPIEN LWAELKKRVR ARRPTNLTQL

301
HQLCQEEWAK IHPTYCGKLV EGYPKRLTQV KQFKGNATKY.

In certain embodiments, including those wherein the Sleeping Beauty transposase is a hyperactive Sleeping Beauty (SB100X) transposase, the Sleeping Beauty transposase enzyme comprises an amino acid sequence at least at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14486)

1
MGKSKEISQD LRKRIVDLHK SGSSLGAISK RLAVPRSSVQ TIVRKYKHHG TTQPSYRSGR

61
RRVLSPRDER TLVRKVQINP RTTAKDLVKM LEETGTKVSI STVKRVLYRH NLKGHSARKK

121
PLLQNRHKKA RLRFATAHGD KDRTFWRNVL WSDETKIELF GHNDHRYVWR KKGEACKPKN

181
TIPTVKHGGG SIMLWGCFAA GGTGALHKID GIMDAVQYVD ILKQHLKTSV RKLKLGRKWV

241
FQHDNDPKHT SKVVAKWLKD NKVKVLEWPS QSPDLNPIEN LWAELKKRVR ARRPTNLTQL

301
HQLCQEEWAK IHPNYCGKLV EGYPKRLTQV KQFKGNATKY.

In certain embodiments of the methods of the disclosure, the transposon is a Helraiser transposon. In certain embodiments of the Helraiser transposon sequence, the transposase is flanked by left and right terminal sequences termed LTS and RTS. In certain embodiments, these sequences terminate with a conserved 5′-TC/CTAG-3′ motif. In certain embodiments, a 19 bp palindromic sequence with the potential to form the hairpin termination structure is located 11 nucleotides upstream of the RTS and comprises the sequence

(SEQ ID NO: 14500)

GTGCACGAATTTCGTGCACCGGGCCACTAG.

In certain embodiments of the methods of the disclosure, and, in particular those embodiments wherein the transposon is a Helraiser transposon, the transposase enzyme is a Helitron transposase enzyme. In certain embodiments, the Helitron transposase enzyme of the disclosure comprises an amino acid sequence comprising:

(SEQ ID NO: 14501)

1
MSKEQLLIQR SSAAERCRRY RQKMSAEQRA SDLERRRRLQ QNVSEEQLLE KRRSEAEKQR

61
RHRQKMSKDQ RAFEVERRRW RRQNMSREQS STSTTNTGRN CLLSKNGVHE DAILEHSCGG

121
MTVRCEFCLS LNFSDEKPSD GKFTRCCSKG KVCPNDIHFP DYPAYLKRLM TNEDSDSKNF

181
MENIRSINSS FAFASMGANI ASPSGYGPYC FRIHGQVYHR TGTLHPSDGV SRKFAQLYIL

241
DTAEATSKRL AMPENQGCSE RLMININNLM HEINELTKSY KMLHEVEKEA QSEAAAKGIA

301
PTEVIMAIKY DRNSDPGRYN SPRVTEVAVI FRNEDGEPPF ERDLLIHCKP DPNNPNATKM

361
KQISILFPTL DAMTYPILFP HGEKGWGTDI ALRLRDNSVI DNNTRQNVRT RVTQMQYYGF

421
HLSVRDTFNP ILNAGKLTQQ FIVDSYSKME ANRINFIKAN QSKLRVEKYS GLMDYLKSRS

481
ENDNVPIGKM IILPSSFEGS PRNMQQRYQD AMAIVTKYGK PDLFITMTCN PKWADITNNL

541
QRWQKVENRP DLVARVFNIK LNALLNDICK FHLFGKVIAK IHVIEFQKRG LPHAHILLIL

601
DSESKLRSED DIDRIVKAEI PDEDQCPRLF QIVKSNMVHG PCGIQNPNSP CMENGKCSKG

661
YPKEFQNATI GNIDGYPKYK RRSGSTMSIG NKVVDNTWIV PYNPYLCLKY NCHINVEVCA

721
SIKSVKYLFK YIYKGHDCAN IQISEKNIIN HDEVQDFIDS RYVSAPEAVW RLFAMRMHDQ

781
SHAITRLAIH LPNDQNLYFH TDDFAEVLDR AKRHNSTLMA WFLLNREDSD ARNYYYWEIP

841
QHYVFNNSLW TKRRKGGNKV LGRLFTVSFR EPERYYLRLL LLHVKGAISF EDLRTVGGVT

901
YDTFHEAAKH RGLLLDDTIW KDTIDDAIIL NMPKQLRQLF AYICVFGCPS AADKLWDENK

961
SHFIEDFCWK LHRREGACVN CEMHALNEIQ EVFTLHGMKC SHFKLPDYPL LMNANTCDQL

1021
YEQQQAEVLI NSLNDEQLAA FQTITSAIED QTVHPKCFFL DGPGGSGKTY LYKVLTHYIR

1081
GRGGTVLPTA STGIAANLLL GGRTFHSQYK LPIPLNETSI SRLDIKSEVA KTIKKAQLLI

1141
IDECTMASSH AINAIDRLLR EIMNLNVAFG GKVLLLGGDF RQCLSIVPHA MRSAIVQTSL

1201
KYCNVWGCFR KLSLKTNMRS EDSAYSEWLV KLGDGKLDSS FHLGMDIIEI PHEMICNGSI

1261
IEATFGNSIS IDNIKNISKR AILCPKNEHV QKLNEEILDI LDGDFHTYLS DDSIDSTDDA

1321
EKENFPIEFL NSITPSGMPC HKLKLKVGAI IMLLRNLNSK WGLCNGTRFI IKRLRPNIIE

1381
AEVLTGSAEG EVVLIPRIDL SPSDTGLPFK LIRRQFPVMP AFAMTINKSQ GQTLDRVGIF

1441
LPEPVFAHGQ LYVAFSRVRR ACDVKVKVVN TSSQGKLVKH SESVFTLNVV YREILE.

In certain embodiments of the methods of the disclosure, the transposon is a Tol2 transposon.

In certain embodiments of the methods of the disclosure, and, in particular those embodiments wherein the transposon is a Tol2 transposon, the transposase enzyme is a Tol2 transposase enzyme. In certain embodiments, the Tol2 transposase enzyme of the disclosure comprises an amino acid sequence comprising:

(SEQ ID NO: 14502)

1
MEEVCDSSAA ASSTVQNQPQ DQEHPWPYLR EFFSLSGVNK DSFKMKCVLC LPLNKEISAF

61
KSSPSNLRKH IERMHPNYLK NYSKLTAQKR KIGTSTHASS SKQLKVDSVF PVKHVSPVTV

121
NKAILRYIIQ GLHPFSTVDL PSFKELISTL QPGISVITRP TLRSKIAEAA LIMKQKVTAA

181
MSEVEWIATT TDCWTARRKS FIGVTAHWIN PGSLERHSAA LACKRLMGSH TFEVLASAMN

241
DIHSEYEIRD KVVCTTTDSG SNFMKAFRVF GVENNDIETE ARRCESDDTD SEGCGEGSDG

301
VEFQDASRVL DQDDGFEFQL PKHQKCACHL LNLVSSVDAQ KALSNEHYKK LYRSVFGKCQ

361
ALWNKSSRSA LAAEAVESES RLQLLRPNQT RWNSTFMAVD RILQICKEAG EGALRNICTS

421
LEVPMFNPAE MLFLTEWANT MRPVAKVLDI LQAETNTQLG WLLPSVHQLS LKLQRLHHSL

481
RYCDPLVDAL QQGIQTRFKH MFEDPEIiAA AILLPKFRTS WTNDETIIKR GMDYIRVHLE

541
PLDHKKELAN SSSDDEDFFA SLKPTTHEAS KELDGYLACV SDTRESLLTF PAICSLSIKT

601
NTPLPASAAC ERLFSTAGLL FSPKRARLDT NNFENQLLLK LNLRFYNFE.

In certain embodiments of the methods of the disclosure, the piggyBac-like transposon comprises an amino acid sequence having at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or any percentage in between of identity to the amino acid sequence of SEQ ID NO: 14487.

In certain embodiments of the methods of the disclosure, a vector comprises the recombinant and non-naturally occurring DNA sequence encoding the transposon. In some embodiments, the vector comprises any form of DNA and wherein the vector comprises at least 100 nucleotides (nts), 500 nts, 1000 nts, 1500 nts, 2000 nts, 2500 nts, 3000 nts, 3500 nts, 4000 nts, 4500 nts, 5000 nts, 6500 nts, 7000 nts, 7500 nts, 8000 nts, 8500 nts, 9000 nts, 9500 nts, 10,000 nts or any number of nucleotides in between. In some embodiments, the vector comprises single-stranded or double-stranded DNA. In some embodiments, the vector comprises circular DNA. In some embodiments, the vector is a plasmid vector. In some embodiments, the vector is a nanoplasmid vector. In some embodiments, the vector is a minicircle. In some embodiments, the vector comprises linear or linearized DNA. In some embodiments, the linear or linearized DNA is produced in vitro. In some embodiments, the linear or linearized DNA is a product of a restriction digest of a circular DNA. In some embodiments, the circular DNA is a plasmid vector, a nanoplasmid vector or a minicircle DNA vector. In some embodiments, the linear or linearized DNA is a product of a polymerase chain reaction (PCR). In some embodiments, the vector is a double-stranded Doggybone™ DNA sequence. In some embodiments, the Doggybone™ DNA sequence is produced by an enzymatic process that solely encodes an antigen expression cassette, comprising antigen, promoter, poly-A tail and telomeric ends.

In certain embodiments of the methods of the disclosure, the immune cell or the immune cell precursor is isolated or derived from a human. In certain embodiments, the immune cell or the immune cell precursor is isolated or derived from a non-human mammal. In certain embodiments, the non-human mammal is a rodent, a rabbit, a cat, a dog, a pig, a horse, a cow, a camel or a primate.

Chimeric antigen receptors (CARs) of the disclosure may comprise (a) an ectodomain comprising an antigen recognition region, (b) a transmembrane domain, and (c) an endodomain comprising at least one costimulatory domain. In certain embodiments, the ectodomain may further comprise a signal peptide. Alternatively, or in addition, in certain embodiments, the ectodomain may further comprise a hinge between the antigen recognition region and the transmembrane domain. In certain embodiments of the CARs of the disclosure, the signal peptide may comprise a sequence encoding a human CD2, CD3δ, CD3ε, CD3γ, CD3ζ, CD4, CD8α, CD19, CD28, 4-1BB or GM-CSFR signal peptide. In certain embodiments of the CARs of the disclosure, the signal peptide may comprise a sequence encoding a human CD8α signal peptide. In certain embodiments, the transmembrane domain may comprise a sequence encoding a human CD2, CD3δ, CD3ε, CD3γ, CD3ζ, CD4, CD8α, CD19, CD28, 4-1BB or GM-CSFR transmembrane domain. In certain embodiments of the CARs of the disclosure, the transmembrane domain may comprise a sequence encoding a human CD8α transmembrane domain. In certain embodiments of the CARs of the disclosure, the endodomain may comprise a human CD3 endodomain. In certain embodiments of the CARs of the disclosure, the at least one costimulatory domain may comprise a human 4-1BB, CD28, CD40, ICOS, MyD88, OX-40 intracellular segment, or any combination thereof. In certain embodiments of the CARs of the disclosure, the at least one costimulatory domain may comprise a CD28 and/or a 4-1BB costimulatory domain. In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD8α, IgG4, and/or CD4 sequence. In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD8α sequence.

In certain embodiments of the methods of the disclosure, the recombinant and non-naturally occurring DNA sequence encoding a transposon further comprises a sequence encoding a chimeric antigen receptor or a portion thereof. The portion of the sequence encoding a chimeric antigen receptor may encode an antigen recognition region. The antigen recognition region may comprise one or more complementarity determining region(s). The antigen recognition region may comprise an antibody, an antibody mimetic, a protein scaffold or a fragment thereof. In certain embodiments, the antibody is a chimeric antibody, a recombinant antibody, a humanized antibody or a human antibody. In certain embodiments, the antibody is affinity-tuned. Nonlimiting examples of antibodies of the disclosure include a single-chain variable fragment (scFv), a VHH, a single domain antibody (sdAB), a small modular immunopharmaceutical (SMIP) molecule, or a nanobody. In certain embodiments, the VHH is camelid. Alternatively, or in addition, in certain embodiments, the VHH is humanized. Nonlimiting examples of antibody fragments of the disclosure include a complementary determining region, a variable region, a heavy chain, a light chain, or any combination thereof. Nonlimiting examples of antibody mimetics of the disclosure include an affibody, an afflilin, an affimer, an affitin, an alphabody, an anticalin, and avimer, a DARPin, a Fynomer, a Kunitz domain peptide, or a monobody. Nonlimiting examples of protein scaffolds of the disclosure include a Centyrin.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 10.0 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 100 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 7.5 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 75 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 6.0 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 60 μg/mL. In certain embodiments, the transposase is a Sleeping Beauty transposase. In certain embodiments, the Sleeping Beauty transposase is a Sleeping Beauty 100X (SB100X) transposase.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 5.0 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 50 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 2.5 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 25 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 1.67 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 16.7 μg/mL. In certain embodiments, the transposase is a Super piggyBac (PB) transposase. In certain embodiments, the piggyBac transposase comprises an amino acid sequence comprising SEQ ID NO: 14487.

In certain embodiments of the methods of the disclosure, the transposase is a piggyBac transposase. In certain embodiments, the piggyBac transposase comprises an amino acid sequence comprising SEQ ID NO: 14487. In certain embodiments, the piggyBac transposase is a hyperactive variant and wherein the hyperactive variant comprises an amino acid substitution at one or more of positions 30, 165, 282 and 538 of SEQ ID NO: 14487. In certain embodiments, the amino acid substitution at position 30 of SEQ ID NO: 14487 is a substitution of a valine (V) for an isoleucine (I) (I30V). In certain embodiments, the amino acid substitution at position 165 of SEQ ID NO: 14487 is a substitution of a serine (S) for a glycine (G) (G165S). In certain embodiments, the amino acid substitution at position 282 of SEQ ID NO: 14487 is a substitution of a valine (V) for a methionine (M) (M282V). In certain embodiments, the amino acid substitution at position 538 of SEQ ID NO: 14487 is a substitution of a lysine (K) for an asparagine (N) (N538K).

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and (b) wherein an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 1.67 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 16.7 μg/mL. In certain embodiments, the transposase is a Super piggyBac (PB) transposase. In certain embodiments, the Super piggyBac (PB) transposase enzyme comprises an amino acid sequence at least 75% identical to:

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 0.55 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 5.5 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 0.19 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 1.9 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 0.10 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 1.0 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 10.0 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 100 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 7.5 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 75 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 6.0 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 60 μg/mL. In certain embodiments, the transposase is a Sleeping Beauty transposase. In certain embodiments, the Sleeping Beauty transposase is a Sleeping Beauty 100X (SB100X) transposase.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 5.0 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 50 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 2.5 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 25 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 1.67 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 16.7 μg/mL. In certain embodiments, the transposase is a Super piggyBac (PB) transposase.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 0.55 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 5.5 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 0.19 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 1.9 μg/mL.

In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 0.1 μg per 100 μL of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 1.0 μg/mL.

The disclosure provides an immune cell modified according to the method of the disclosure. The immune cell may be a T-lymphocyte, a Natural Killer (NK) cell, a Cytokine-induced Killer (CIK) cell or a Natural Killer T (NKT) cell. The immune cell may be further modified by a second gene editing tool, including, but not limited to those gene editing tools comprising an endonuclease operably-linked to either a Cas9 or a TALE sequence. In certain embodiments of the second gene editing tool, the endonuclease is operably-linked to either a Cas9 or a TALE sequence covalently. In certain embodiments of the second gene editing tool, the endonuclease is operably-linked to either a Cas9 or a TALE sequence non-covalently. In certain embodiments, the endonuclease comprises a Clo051 domain. In certain embodiments, Clo051 domain comprises a sequence of

(SEQ ID NO: 14503)

EGIKSNISLLKDELRGQISHISHEYLSLIDLAFDSKQNRLFEMKVLEL

LVNEYGFKGRHLGGSRKPDGIVYSTTLEDNFGIIVDTKAYSEGYSLPI

SQADEMERYVRENSNRDEEVNPNKWWENFSEEVKKYYFVFISGSFKGK

FEEQLRRLSMTTGVNGSAVNVVNLLLGAEKIRSGEMTIEELERAMFNN

SEFILKY.

In certain embodiments, the Cas9 is an inactivated Cas9 (dCas9). In certain embodiments, the inactivated Cas9 is isolated or derived from Staphylococcus aureus and comprises D10A and N580A within the catalytic site. In certain embodiments, the Cas9 is a small and inactivated Cas9 (dSaCas9). In certain embodiments, the dSaCas9 comprises the amino acid sequence of

(SEQ ID NO: 14497)

1
MKRNYILGLA IGITSVGYGI IDYETRDVID AGVRLFKEAN VENNEGRRSK RGARRLKRRR

61
RHRIQRVKKL LFDYNLLTDH SELSGINPYE ARVKGLSQKL SEEEFSAALL HLAKRRGVHN

121
VNEVEEDTGN ELSTKEQISR NSKALEEKYV AELQLERLKK DGEVRGSINR FKTSDYVKEA

181
KQLLKVQKAY HQLDQSFIDT YIDLLETRRT YYEGPGEGSP FGWKDIKEWY EMLMGHCTYF

241
PEELRSVKYA YNADLYNALN DLNNLVITRD ENEKLEYYEK FQIIENVFKQ KKKPTLKQIA

301
KEILVNEEDI KGYRVTSTGK PEFTNLKVYH DIKDITARKE IIENAELLDQ IAKILTIYQS

361
SEDIQEELTN LNSELTQEEI EQISNLKGYT GTHNLSLKAI NLILDELWHT NDNQIAIFNR

421
LKLVPKKVDL SQQKEIPTTL VDDFILSPVV KRSFIQSIKV INAIIKKYGL PNDIIIELAR

481
EKNSKDAQKM INEMQKRNRQ TNERIEEIIR TTGKENAKYL IEKIKLHDMQ EGKCLYSLEA

541
IPLEDLLNNP FNYEVDHIIP RSVSFDNSFN NKVLVKQEEA SKKGNRTPFQ YLSSSDSKIS

601
YETFKKHILN LAKGKGRISK TKKEYLLEER DINRFSVQKD FINRNLVDTR YATRGLMNLL

661
RSYFRVNNLD VKVKSINGGF TSFLRRKWKF KKERNKGYKH HAEDALIIAN ADFIFKEWKK

721
LDKAKKVMEN QMFEEKQAES MPEIETEQEY KEIFITPHQI KHIKDFKDYK YSHRVDKKPN

781
RELINDTLYS TRKDDKGNTL IVNNLNGLYD KDNDKLKKLI NKSPEKLLMY HHDPQTYQKL

841
KLIMEQYGDE KNPLYKYYEE TGNYLTKYSK KDNGPVIKKI KYYGNKLNAH LDITDDYPNS

901
RNKVVKLSLK PYRFDVYLDN GVYKFVTVKN LDVIKKENYY EVNSKCYEEA KKLKKISNQA

961
EFIASFYNND LIKINGELYR VIGVNNDLLN RIEVNMIDIT YREYLENMND KRPPRIIKTI

1021
ASKTQSIKKY STDILGNLYE VKSKKHPQII KKG.

In certain embodiments, the Cas9 is an inactivated Cas9 (dCas9). In certain embodiments, the inactivated Cas9 (dCas9) is isolated or derived from Staphylococcus pyogenes and comprises D10A and H840A within the catalytic site. In certain embodiments, the dCas9 comprises the amino acid sequence of:

(SEQ ID NO: 14498)

1
XDKKYSIGLA IGTNSVGWAV ITDEYKVPSK KFKVLGNTDR HSIKKNLIGA LLFDSGETAE

61
ATRLKRTARR RYTRRKNRIC YLQEIFSNEM AKVDDSFFHR LEESFLVEED KKHERHPIFG

121
NIVDEVAYHE KYPTIYHLRK KLVDSTDKAD LRLIYLALAH MIKFRGHFLI EGDLNPDNSD

181
VDKLFIQLVQ TYNQLFEENP INASGVDAKA ILSARLSKSR RLENLIAQLP GEKKNGLFGN

241
LIALSLGLTP NFKSNFDLAE DAKLQLSKDT YDDDLDNLLA QIGDQYADLF LAAKNLSDAI

301
LLSDILRVNT EITKAPLSAS MIKRYDEHHQ DLTLLKALVR QQLPEKYKEI FFDQSKNGYA

361
GYIDGGASQE EFYKFIKPIL EKMDGTEELL VKLNREDLLR KQRTFDNGSI PHQIHLGELH

421
AILRRQEDFY PFLKDNREKI EKILTFRIPY YVGPLARGNS RFAWMTRKSE ETITPWNFEE

481
VVDKGASAQS FIERMTNFDK NLPNEKVLPK HSLLYEYFTV YNELTKVKYV TEGMRKPAFL

541
SGEQKKAIVD LLFKTNRKVT VKQLKEDYFK KIECFDSVEI SGVEDRFNAS LGTYHDLLKI

601
IKDKDFLDNE ENEDILEDIV LTLTLFEDRE MIEERLKTYA HLFDDKVMKQ LKRRRYTGWG

661
RLSRKLINGI RDKQSGKTIL DFLKSDGFAN RNFMQLIHDD SLTFKEDIQK AQVSGQGDSL

721
HEHIANLAGS PAIKKGILQT VKVVDELVKV MGRHKPENIV IEMARENQTT QKGQKNSRER

781
MKRIEEGIKE LGSQILKEHP VENTQLQNEK LYLYYLQNGR DMYVDQELDI NRLSDYDVDA

841
IVPQSFLKDD SIDNKVLTRS DKNRGKSDNV PSEEVVKKMK NYWRQLLNAK LITQRKFDNL

901
TKAERGGLSE LDKAGFIKRQ LVETRQITKH VAQILDSRMN TKYDENDKLI REVKVITLKS

961
KLVSDFRKDF QFYKVREINN YHHAHDAYLN AVVGTALIKK YPKLESEFVY GDYKVYDVRK

1021
MIAKSEQEIG KATAKYFFYS NIMNFFKTEI TLANGEIRKR PLIETNGETG EIVWDKGRDF

1081
ATVRKVLSMP QVNIVKKTEV QTGGFSKESI LPKRNSDKLI ARKKDWDPKK YGGFDSPTVA

1141
YSVLVVAKVE KGKSKKLKSV KELLGITIME RSSFEKNPID FLEAKGYKEV KKDLIIKLPK

1201
YSLFELENGR KRMLASAGEL QKGNELALPS KYVNFLYLAS HYEKLKGSPE DNEQKQLFVE

1261
QHKHYLDEII EQISEFSKRV ILADANLDKV LSAYNKHRDK PIREQAENII HLFTLTNLGA

1321
PAAFKYFDTT IDRKRYTSTK EVLDATLIHQ SITGLYETRI DLSQLGGD.

(SEQ ID NO: 14499)

1
MDKKYSIGLA IGTNSVGWAV ITDEYKVPSK KFKVLGNTDR HSIKKNLIGA LLFDSGETAE

61
ATRLKRTARR RYTRRKNRIC YLQEIFSNEM AKVDDSFFHR LEESFLVEED KKHERHPIFG

121
NIVDEVAYHE KYPTIYHLRK KLVDSTDKAD LRLIYLALAH MIKFRGHFLI EGDLNPDNSD

181
VDKLFIQLVQ TYNQLFEENP INASGVDAKA ILSARLSKSR RLENLIAQLP GEKKNGLFGN

241
LIALSLGLTP NFKSNFDLAE DAKLQLSKDT YDDDLDNLLA QIGDQYADLF LAAKNLSDAI

301
LLSDILRVNT EITKAPLSAS MIKRYDEHHQ DLTLLKALVR QQLPEKYKEI FFDQSKNGYA

361
GYIDGGASQE EFYKFIKPIL EKMDGTEELL VKLNREDLLR KQRTFDNGSI PHQIHLGELH

421
AILRRQEDFY PFLKDNREKI EKILTFRIPY YVGPLARGNS RFAWMTRKSE ETITPWNFEE

481
VVDKGASAQS FIERMTNFDK NLPNEKVLPK HSLLYEYFTV YNELTKVKYV TEGMRKPAFL

541
SGEQKKAIVD LLFKTNRKVT VKQLKEDYFK KIECFDSVEI SGVEDRFNAS LGTYHDLLKI

601
IKDKDFLDNE ENEDILEDIV LTLTLFEDRE MIEERLKTYA HLFDDKVMKQ LKRRRYTGWG

661
RLSRKLINGI RDKQSGKTIL DFLKSDGFAN RNFMQLIHDD SLTFKEDIQK AQVSGQGDSL

721
HEHIANLAGS PAIKKGILQT VKVVDELVKV MGRHKPENIV IEMARENQTT QKGQKNSRER

781
MKRIEEGIKE LGSQILKEHP VENTQLQNEK LYLYYLQNGR DMYVDQELDI NRLSDYDVDA

841
IVPQSFLKDD SIDNKVLTRS DKNRGKSDNV PSEEVVKKMK NYWRQLLNAK LITQRKFDNL

901
TKAERGGLSE LDKAGFIKRQ LVETRQITKH VAQILDSRMN TKYDENDKLI REVKVITLKS

961
KLVSDFRKDF QFYKVREINN YHHAHDAYLN AVVGTALIKK YPKLESEFVY GDYKVYDVRK

1021
MIAKSEQEIG KATAKYFFYS NIMNFFKTEI TLANGEIRKR PLIETNGETG EIVWDKGRDF

1081
ATVRKVLSMP QVNIVKKTEV QTGGFSKESI LPKRNSDKLI ARKKDWDPKK YGGFDSPTVA

1141
YSVLVVAKVE KGKSKKLKSV KELLGITIME RSSFEKNPID FLEAKGYKEV KKDLIIKLPK

1201
YSLFELENGR KRMLASAGEL QKGNELALPS KYVNFLYLAS HYEKLKGSPE DNEQKQLFVE

1261
QHKHYLDEII EQISEFSKRV ILADANLDKV LSAYNKHRDK PIREQAENII HLFTLTNLGA

1321
PAAFKYFDTT IDRKRYTSTK EVLDATLIHQ SITGLYETRI DLSQLGGD.

The disclosure provides an immune cell modified according to the method of the disclosure. The immune cell may be a T-lymphocyte, a Natural Killer (NK) cell, a Cytokine-induced Killer (CIK) cell or a Natural Killer T (NKT) cell. The immune cell may be further modified by a second gene editing tool, including, but not limited to those gene editing tools comprising an endonuclease operably-linked to either a Cas9 or a TALE sequence. Alternatively or in addition, the second gene editing tool may include an excision-only piggyBac transposase to re-excise the inserted sequences or any portion thereof. For example, the excision-only piggyBac transposase may be used to “re-excise” the transposon.

In certain embodiments, the transposon is a piggyBac transposon. In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac transposon, the transposase is a piggyBac™ or a Super piggyBac™ (SPB) transposase. In certain embodiments, and, in particular, those embodiments wherein the transposase is a Super piggyBac™ (SPB) transposase, the sequence encoding the transposase is an mRNA sequence.

(SEQ ID NO: 14487)

1
MGSSLDDEHI LSALLQSDDE LVGEDSDSEI SDHVSEDDVQ

SDTEEAFIDE VHEVQPTSSG

61
SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST

SKSTRRSRVS ALNIVRSQRG

121
PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR

ESMTGATFRD TNEDEIYAFF

181
GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL

IRCLRMDDKS IRPTLRENDV

241
FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF

RMYIPNKPSK YGIKILMMCD

301
SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC

RNITCDNWFT SIPLAKNLLQ

361
EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP

LTLVSYKPKP AKMVYLLSSC

421
DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR

KTNRWPMALL YGMINIACIN

481
SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE

APTLKRYLRD NISNILPNEV

541
PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV

ICREHNIDMC QSCF.

(SEQ ID NO: 14484)

1
MGSSLDDEHI LSALLQSDDE LVGEDSDSEV SDHVSEDDVQ

SDTEEAFIDE VHEVQPTSSG

61
SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST

SKSTRRSRVS ALNIVRSQRG

121
PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR

ESMTSATFRD TNEDEIYAFF

181
GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL

IRCLRMDDKS IRPTLRENDV

241
FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF

RVYIPNKPSK YGIKILMMCD

301
SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC

RNITCDNWFT SIPLAKNLLQ

361
EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP

LTLVSYKPKP AKMVYLLSSC

421
DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR

KTNRWPMALL YGMINIACIN

481
SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE

APTLKRYLRD NISNILPKEV

541
PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV

ICREHNIDMC QSCF.

The disclosure provides a culture media for enhancing viability of a modified immune cell comprising IL-2, IL-21, IL-7, IL-15 or any combination thereof. The modified immune cell may be a T-lymphocyte, a Natural Killer (NK) cell, a Cytokine-induced Killer (CIK) cell or a Natural Killer T (NKT) cell. In some embodiments, the modified immune cell is a T-lymphocyte. In some embodiments, the T-lymphocyte is an early memory T-cell. In some embodiments, the T-lymphocyte is a stem cell-like T-cell. In some embodiments, the T-lymphocyte is a stem memory T cell (T_SCM). In some embodiments, the T-lymphocyte is a central memory T cell (T_CM). The modified immune cell may contain one or more exogenous DNA sequences. The modified immune cell may contain one or more exogenous RNA sequences. The modified immune cell may have been electroporated or nucleofected.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a series of graphs depicting transfection efficiency and cell viability following plasmid DNA nucleofection in primary human T lymphocytes.

FIG. 2 is a series of graphs depicting DNA cytotoxicity to T cells.

FIG. 3 is a series of graphs showing that DNA-mediated cytotoxicity in T cells is dose dependent.

FIG. 4 is a series of graphs showing that extracellular plasmid DNA is not cytotoxic.

FIG. 5 is a series of graphs depicting efficient transposition using SPB mRNA in Jurkat cells.

FIG. 6 is a series of graphs depicting efficient transposition in T lymphocytes using SPB mRNA.

FIG. 7 is a series of graphs depicting efficient delivery of linearized DNA transposon products.

FIG. 8 is a series of graphs showing that addition of that IL-7 and IL-15 and immediate stimulation of T cells post-nucleofection enhances cell viability.

FIG. 9 is a series of graphs showing that IL-7 and IL-15 rescue T cells from DNA mediated toxicity

FIG. 10 is a series of graphs showing that immediate stimulation of T cells post-nucleofection enhances cell viability.

FIG. 11A-C is a series of graphs depicting T cell transposition with varying amounts of DNA. Primary human pan T cells were nucleofected with varying amounts of DNA using piggyBac™. T cells were nucleofected with the indicated amounts of transposon and 5 μg SPB mRNA. Cells were then stimulated on day 2 post-nucleofection through CD3 and CD28. As expected, T cells nucleofected with high amounts of DNA exhibited high episomal expression at day 1 post nucleofection whereas almost no episomal expression was observed at low DNA doses. In contrast, following expansion at day 21 post nucleofection the greatest percentage of transgene positive cells were observed in lower DNA amounts peaking at 1.67 μg for this transposon. (A) Flow analysis for transgene positive cells at day 1 and 21. (B) Percentage of transgene positive T cells. (C) Percentage of viable T cells at day 1 and 21. For all graphs shown in this figure, the Y-axis ranges from 0 to 100% in increments of 20% and the X-axis ranges from 0 to 10⁵by powers of 10.

FIG. 12A-B is a series of graphs depicting T cell transposition with low DNA amounts using the Sleeping Beauty™ 100X (SB100X) transposase. Primary human pan T cells were nucleofected with GFP plasmids encoding either the piggyBac™ (PB) or Sleeping Beauty™ (SB) ITRs. (A) Cells were nucleofected with the indicated amounts of SB transposon and 1 μg SB transposase mRNA. (B) Cells were nucleofected with the indicated amounts of SB transposase and 0.75 μg SB transposon. Flow analysis was performed on day 14 post nucleofection for all samples. For all graphs shown in this figure, the Y-axis ranges from 0 to 250K in increments of 50K and the X-axis ranges from 0 to 10⁵by powers of 10.

FIG. 13A is a series of plots depicting T cells transposed with a plasmid containing a sequence encoding a transposon comprising a sequence encoding an inducible caspase polypeptide (a safety switch, “iC9”), a CARTyrin (anti-BCMA), and a selectable marker. Left-hand plots depict live T cells exposed to transposase in the absence of the plasmid. Right-hand plots depict live T cells exposed to transposase in the presence of the plasmid. Cells were exposed to either a hyperactive transposase (the “Super piggyBac”) or a wild type piggyBac transposase.

FIG. 13B is a series of plots depicting T cells transposed with a plasmid containing a sequence encoding a green fluorescent protein (GFP). Left-hand plots depict live T cells exposed to transposase in the absence of the plasmid. Right-hand plots depict live T cells exposed to transposase in the presence of the plasmid. Cells were exposed to either a hyperactive transposase (the “Super piggyBac”) or a wild type piggyBac transposase.

FIG. 13C is a table depicting the percent of transformed T cells resulting from transposition with WT versus hyperactive piggyBac transposase. T cells contacted with the hyperactive piggyBac transposase (the Super piggyBac transposase) were transformed at a rate 4-fold greater than WT transposase.

FIG. 13D is a graph depicting the percent of transformed T cells resulting from transposition with WT versus hyperactive piggyBac transposase 5 days after nucleofection. T cells contacted with the hyperactive piggyBac transposase (the Super piggyBac transposase) were transformed at a rate far greater than WT transposase.

FIG. 14 is a graph depicting transposition in natural killer (NK) cells. Transposition of non-activated NK cells derived from CD3-depleted leukopheresis (containing CD14/CD19/CD56+ cells) is shown. Cells were electroporated (EP) with plasmid piggyBac transposon DNA encoding GFP and mRNA encoding super piggyBac. The program from Lonza 4D nucleofector or BTX ECM 830 (500V, 700 usec pulse length, 0.2 mm electrode gap, one pulse) is indicated on the X-axis. Transposed cells were co-cultured (stimulated) at day 2 with artificial antigen presenting cells (aAPCs). Fluorescent activated cell sorting (FACS) analysis of percent GFP positive cells at day 7 post-EP (day 5 post-stim) is indicated on the Y-axis with gray bars. Percent viability as shown by percent 7-Aminoactinomycin D (7AAD)-negative cells at day 2 post-EP is indicated on the Y-axis with gray bars.

FIG. 15A-B are a series of 10 FACs plots (FIG. 15A) and a graph (FIG. 15B) showing transposon titration for transposition in natural killer (NK) cells. Transposition of non-activated NK cells from CD3-depleted leukopheresis (containing CD14/CD19/CD56+ cells) is shown. Cells were electroporated with a plasmid piggyBac transposon encoding GFP at amounts ranging from 0 to 10 ug of DNA and 5 ug mRNA encoding Super piggyBac using the indicated Maxcyte electroporator program. Transposed cells were stimulated at day 2 with artificial antigen presenting cells (aAPCs). FIG. 15A FACs plots top row shows CD56+(y-axis) versus GFP+(x-axis) expression, while the bottom row shows 7AAD (y-axis) versus forward scatter (FSC, x-axis). FIG. 15B is a bar graph analysis of the percentage of GFP+ cells of CD56+ cells at day 6 post-electroporation (EP) and day 4 post-stimulation (black bars), and the percent viability as shown by 7AAD-negative cells at day 2 post EP (gray bars).

FIG. 16A-B are a series of 7 FACs plots (FIG. 16A) and a graph (FIG. 16B) showing dose-dependent DNA-mediated cytotoxicity in NK cells. FACS analysis of live cells (7AAD-negative/FSC) at day 2 post-EP using the Lonza 4D Nucleofector program DN-100 are shown (FIG. 16A). FACS plots (FIG. 16A) are quantified in a graph (FIG. 16B). 5E6 cells per EP were electroporated in 100 uL P3 buffer in cuvettes. Cells were electroporated with no DNA (Mock) or varying amounts of piggyBac GFP transposon co-delivered with 5 ug Super piggyBac mRNA.

FIG. 17 is a series of 5 graphs showing the in vitro differentiation of piggyBac modified hematopoietic stem and precursor cells (HSPCs) into B cells. Human CD34+ HSPCs were electroporated with mRNA encoding Super piggyBac along with a piggyBac transposon encoding GFP. After electroporation, HSPCs were primed for B cell differentiation in presence of human IL-3, Flt3L, TPO, SCF, and G-CSF for 5 days. On day 6, cells were transferred to a layer of MS-5 feeder cells and fed bi-weekly, along with transfer to a fresh layer of feeders once per week. On day 34 of the in vitro differentiation process, CD19+ B cells were generated and detectable in the culture. Top row: FACs plots showing CD19 (y-axis) and CD34 (x-axis) in, from left to right, human primary bone marrow cells, at day 6 of in vitro differentiation, and at day 34 of in vitro differentiation. Bottom row: graphs depicting GFP expression in the indicated boxed populations of cells from the FACs plots in the top row at days 6 and 34 of in vitro differentiation.

FIG. 18 is a schematic depiction of the Csy4-T2A-Clo051-G4Slinker-dCas9 construct map.

FIG. 19 is a schematic depiction of the pRT1-Clo051-dCas9 Double NLS construct map.

DETAILED DESCRIPTION

Disclosed are compositions and methods for the ex-vivo genetic modification of an immune cell or a precursor thereof comprising delivering to the immune cell or immune precursor cell, (a) a nucleic acid or amino acid sequence comprising a sequence encoding a transposase enzyme and (b) a recombinant and non-naturally occurring DNA sequence comprising a DNA sequence encoding a transposon. In certain embodiments, the method further comprises the step of stimulating the immune cell or immune precursor cell with one or more cytokine(s).

Immune and Immune Precursor Cells

In certain embodiments, immune cells of the disclosure comprise lymphoid progenitor cells, natural killer (NK) cells, T lymphocytes (T-cell), stem memory T cells (T_SCMcells), Stem cell-like T cells, B lymphocytes (B-cells), myeloid progenitor cells, neutrophils, basophils, eosinophils, monocytes, macrophages, platelets, erythrocytes, red blood cells (RBCs), megakaryocytes or osteoclasts.

In certain embodiments, immune precursor cells comprise any cells which can differentiate into one or more types of immune cells. In certain embodiments, immune precursor cells comprise multipotent stem cells that can self renew and develop into immune cells. In certain embodiments, immune precursor cells comprise hematopoietic stem cells (HSCs) or descendants thereof. In certain embodiments, immune precursor cells comprise precursor cells that can develop into immune cells. In certain embodiments, the immune precursor cells comprise hematopoietic progenitor cells (HPCs).

Hematopoietic Stem Cells (HSCs)

Hematopoietic stem cells (HSCs) are multipotent, self-renewing cells. All differentiated blood cells from the lymphoid and myeloid lineages arise from HSCs. HSCs can be found in adult bone marrow, peripheral blood, mobilized peripheral blood, peritoneal dialysis effluent and umbilical cord blood.

HSCs of the disclosure may be isolated or derived from a primary or cultured stem cell. HSCs of the disclosure may be isolated or derived from an embryonic stem cell, a multipotent stem cell, a pluripotent stem cell, an adult stem cell, or an induced pluripotent stem cell (iPSC).

Immune precursor cells of the disclosure may comprise an HSC or an HSC descendent cell. Exemplary HSC descendent cells of the disclosure include, but are not limited to, multipotent stem cells, lymphoid progenitor cells, natural killer (NK) cells, T lymphocyte cells (T-cells), B lymphocyte cells (B-cells), myeloid progenitor cells, neutrophils, basophils, eosinophils, monocytes, and macrophages.

HSCs produced by the methods of the disclosure may retain features of “primitive” stem cells that, while isolated or derived from an adult stem cell and while committed to a single lineage, share characteristics of embryonic stem cells. For example, the “primitive” HSCs produced by the methods of the disclosure retain their “stemness” following division and do not differentiate. Consequently, as an adoptive cell therapy, the “primitive” HSCs produced by the methods of the disclosure not only replenish their numbers, but expand in vivo. “Primitive” HSCs produced by the methods of the disclosure may be therapeutically-effective when administered as a single dose. In some embodiments, primitive HSCs of the disclosure are CD34+. In some embodiments, primitive HSCs of the disclosure are CD34+ and CD38−. In some embodiments, primitive HSCs of the disclosure are CD34+, CD38− and CD90+. In some embodiments, primitive HSCs of the disclosure are CD34+, CD38−, CD90+ and CD45RA−. In some embodiments, primitive HSCs of the disclosure are CD34+, CD38−, CD90+, CD45RA−, and CD49f+. In some embodiments, the most primitive HSCs of the disclosure are CD34+, CD38−, CD90+, CD45RA−, and CD49f+.

In some embodiments of the disclosure, primitive HSCs, HSCs, and/or HSC descendent cells may be modified according to the methods of the disclosure to express an exogenous sequence (e.g. a chimeric antigen receptor or therapeutic protein). In some embodiments of the disclosure, modified primitive HSCs, modified HSCs, and/or modified HSC descendent cells may be forward differentiated to produce a modified immune cell including, but not limited to, a modified T cell, a modified natural killer cell and/or a modified B-cell of the disclosure.

T Cells

Modified T cells of the disclosure may be derived from modified hematopoietic stem and progenitor cells (HSPCs) or modified HSCs.

Unlike traditional biologics and chemotherapeutics, modified-T cells of the disclosure possess the capacity to rapidly reproduce upon antigen recognition, thereby potentially obviating the need for repeat treatments. To achieve this, in some embodiments, modified-T cells of the disclosure not only drive an initial response, but also persist in the patient as a stable population of viable memory T cells to prevent potential relapses. Alternatively, in some embodiments, when it is not desired, modified-T cells of the disclosure do not persist in the patient.

Intensive efforts have been focused on the development of antigen receptor molecules that do not cause T cell exhaustion through antigen-independent (tonic) signaling, as well as of a modified-T cell product containing early memory T cells, especially stem cell memory (T_SCM) or stem cell-like T cells. Stem cell-like modified-T cells of the disclosure exhibit the greatest capacity for self-renewal and multipotent capacity to derive central memory (T_CM) T cells or T_CMlike cells, effector memory (T_EM) and effector T cells (T_E), thereby producing better tumor eradication and long-term modified-T cell engraftment. A linear pathway of differentiation may be responsible for generating these cells: Naïve T cells (T_N)>T_SCM>T_CM>T_EM>T_E>T_TE, whereby T_Nis the parent precursor cell that directly gives rise to T_SCM, which then, in turn, directly gives rise to T_CM, etc. Compositions of T cells of the disclosure may comprise one or more of each parental T cell subset with T_SCMcells being the most abundant (e.g. T_SCM>T_CM>T_EM>T_E>T_TE).

In some embodiments of the methods of the disclosure, the immune cell precursor is differentiated into or is capable of differentiating into an early memory T cell, a stem cell like T-cell, a Naïve T cells (T_N), a T_SCM, a T_CM, a T_EM, a T_E, or a T_TE. In some embodiments, the immune cell precursor is a primitive HSC, an HSC, or a HSC descendent cell of the disclosure.

In some embodiments of the methods of the disclosure, the immune cell is an early memory T cell, a stem cell like T-cell, a Naïve T cells (T_N), a T_SCM, a T_CM, a T_EM, a T_E, or a T_TE.

In some embodiments of the methods of the disclosure, the immune cell is an early memory T cell.

In some embodiments of the methods of the disclosure, the immune cell is a stem cell like T-cell.

In some embodiments of the methods of the disclosure, the immune cell is a T_SCM.

In some embodiments of the methods of the disclosure, the immune cell is a T_CM.

In some embodiments of the methods of the disclosure, the methods modify and/or the methods produce a plurality of modified T cells, wherein at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of modified T cells expresses one or more cell-surface marker(s) of an early memory T cell. In certain embodiments, the plurality of modified early memory T cells comprises at least one modified stem cell-like T cell. In certain embodiments, the plurality of modified early memory T cells comprises at least one modified T_SCM. In certain embodiments, the plurality of modified early memory T cells comprises at least one modified T_CM.

In some embodiments of the methods of the disclosure, the methods modify and/or the methods produce a plurality of modified T cells, wherein at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of modified T cells expresses one or more cell-surface marker(s) of a stem cell-like T cell. In certain embodiments, the plurality of modified stem cell-like T cells comprises at least one modified T_SCM. In certain embodiments, the plurality of modified stem cell-like T cells comprises at least one modified T_CM.

In some embodiments of the methods of the disclosure, the methods modify and/or the methods produce a plurality of modified T cells, wherein at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of modified T cells expresses one or more cell-surface marker(s) of a stem memory T cell (T_SCM). In certain embodiments, the cell-surface markers comprise CD62L and CD45RA. In certain embodiments, the cell-surface markers comprise one or more of CD62L, CD45RA, CD28, CCR7, CD127, CD45RO, CD95, CD95 and IL-2Rβ. In certain embodiments, the cell-surface markers comprise one or more of CD45RA, CD95, CCR7, and CD62L.

In some embodiments of the methods of the disclosure, a buffer comprises the immune cell or precursor thereof. The buffer maintains or enhances a level of cell viability and/or a stem-like phenotype of the immune cell or precursor thereof, including T-cells. In certain embodiments, the buffer maintains or enhances a level of cell viability and/or a stem-like phenotype of the primary human T cells prior to the nucleofection. In certain embodiments, the buffer maintains or enhances a level of cell viability and/or a stem-like phenotype of the primary human T cells during the nucleofection. In certain embodiments, the buffer maintains or enhances a level of cell viability and/or a stem-like phenotype of the primary human T cells following the nucleofection. In certain embodiments, the buffer comprises one or more of KCl, MgCl₂, ClNa, Glucose and Ca (NO₃)₂in any absolute or relative abundance or concentration, and, optionally, the buffer further comprises a supplement selected from the group consisting of HEPES, Tris/HCl, and a phosphate buffer. In certain embodiments, the buffer comprises 5 mM KCl, 15 mM MgCl₂, 90 mM ClNa, 10 mM Glucose and 0.4 mM Ca(NO₃)₂. In certain embodiments, the buffer comprises 5 mM KCl, 15 mM MgCl₂, 90 mM ClNa, 10 mM Glucose and 0.4 mM Ca(NO₃)₂and a supplement comprising 20 mM HEPES and 75 mM Tris/HCl. In certain embodiments, the buffer comprises 5 mM KCl, 15 mM MgCl₂, 90 mM ClNa, 10 mM Glucose and 0.4 mM Ca(NO₃)₂and a supplement comprising 40 mM Na₂HPO₄/NaH₂PO₄at pH 7.2. In certain embodiments, the composition comprising primary human T cells comprises 100 μl of the buffer and between 5×10⁶and 25×10⁶cells. In certain embodiments, the composition comprises a scalable ratio of 250e6 primary human T cells per milliliter of buffer or other media during the introduction step.

In some embodiments of the methods of the disclosure, the introducing step may comprise delivery of transposon and/or transposase by a method other than electroporation or nucleofection. In some embodiments, a composition comprises a scalable ratio of 250e6 primary human T cells per milliliter of buffer or other media during the introduction step.

In some embodiments of the methods of the disclosure, the introducing step comprises one or more of topical delivery, adsorption, absorption, electroporation, spin-fection, co-culture, transfection, mechanical delivery, sonic delivery, vibrational delivery, magnetofection or by nanoparticle-mediated delivery.

In some embodiments of the methods of the disclosure, the introducing step comprises liposomal transfection, calcium phosphate transfection, fugene transfection, and dendrimer-mediated transfection.

In some embodiments of the methods of the disclosure, the introducing step comprises mechanical transfection comprises cell squeezing, cell bombardment, or gene gun techniques.

In some embodiments of the methods of the disclosure, the introducing step comprises nanoparticle-mediated transfection comprises liposomal delivery, delivery by micelles, and delivery by polymerosomes.

In some embodiments of the methods of the disclosure, the methods comprise contacting an immune cell of the disclosure, including a T cell of the disclosure, and a T-cell expansion composition. In some embodiments of the methods of the disclosure, the step of introducing a transposon and/or transposase of the disclosure into an immune cell of the disclosure may further comprise contacting the immune cell and a T-cell expansion composition. In some embodiments, including those in which the introducing step of the methods comprises an electroporation or a nucleofection step, the electroporation or a nucleofection step may be performed with the immune cell contacting T-cell expansion composition of the disclosure.

In some embodiments of the methods of the disclosure, the T-cell expansion composition comprises, consists essentially of or consists of phosphorus; one or more of an octanoic acid, a palmitic acid, a linoleic acid, and an oleic acid; a sterol; and an alkane.

In certain embodiments of the methods of producing a modified T cell of the disclosure, the expansion supplement comprises one or more cytokine(s). The one or more cytokine(s) may comprise any cytokine, including but not limited to, lymphokines. Exemplary lympokines include, but are not limited to, interleukin-2 (IL-2), interleukin-3 (IL-3), interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-6 (IL-6), interleukin-7 (IL-7), interleukin-15 (IL-15), interleukin-21 (IL-21), granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma (INFγ). The one or more cytokine(s) may comprise IL-2.

In some embodiments of the methods of the disclosure, the T-cell expansion composition comprises human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, and an expansion supplement. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid, nicotinamide, 2,4,7,9-tetramethyl-5-decyn-4,7-diol (TMDD), diisopropyl adipate (DIPA), n-butyl-benzenesulfonamide, 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester, palmitic acid, linoleic acid, oleic acid, stearic acid hydrazide, oleamide, a sterol and an alkane. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid, palmitic acid, linoleic acid, oleic acid and a sterol. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid at a concentration of between 0.9 mg/kg to 90 mg/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; oleic acid at a concentration of 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; and a sterol at a concentration of about 0.1 mg/kg to 10 mg/kg, inclusive of the endpoints. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid at a concentration of about 9 mg/kg, palmitic acid at a concentration of about 2 mg/kg, linoleic acid at a concentration of about 2 mg/kg, oleic acid at a concentration of about 2 mg/kg and a sterol at a concentration of about 1 mg/kg. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid at a concentration of between 6.4 μmol/kg and 640 μmol/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.7 μmol/kg and 70 μmol/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.75 μmol/kg and 75 μmol/kg, inclusive of the endpoints; oleic acid at a concentration of between 0.75 μmol/kg and 75 μmol/kg, inclusive of the endpoints; and a sterol at a concentration of between 0.25 μmol/kg and 25 μmol/kg, inclusive of the endpoints. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid at a concentration of about 64 μmol/kg, palmitic acid at a concentration of about 7 μmol/kg, linoleic acid at a concentration of about 7.5 μmol/kg, oleic acid at a concentration of about 7.5 μmol/kg and a sterol at a concentration of about 2.5 μmol/kg.

In certain embodiments, the T-cell expansion composition comprises one or more of human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, and an expansion supplement to produce a plurality of expanded modified T-cells, wherein at least 2% of the plurality of modified T-cells expresses one or more cell-surface marker(s) of an early memory T cell, a stem cell-like T cell, a stem memory T cell (T_SCM) and/or a central memory T cell (T_CM). In certain embodiments, the T-cell expansion composition comprises or further comprises one or more of octanoic acid, nicotinamide, 2,4,7,9-tetramethyl-5-decyn-4,7-diol (TMDD), diisopropyl adipate (DIPA), n-butyl-benzenesulfonamide, 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester, palmitic acid, linoleic acid, oleic acid, stearic acid hydrazide, oleamide, a sterol and an alkane. In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid, palmitic acid, linoleic acid, oleic acid and a sterol (e.g. cholesterol). In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of between 0.9 mg/kg to 90 mg/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; oleic acid at a concentration of 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; and a sterol at a concentration of about 0.1 mg/kg to 10 mg/kg, inclusive of the endpoints (wherein mg/kg=parts per million). In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of about 9 mg/kg, palmitic acid at a concentration of about 2 mg/kg, linoleic acid at a concentration of about 2 mg/kg, oleic acid at a concentration of about 2 mg/kg, and a sterol at a concentration of about 1 mg/kg (wherein mg/kg=parts per million). In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of 9.19 mg/kg, palmitic acid at a concentration of 1.86 mg/kg, linoleic acid at a concentration of about 2.12 mg/kg, oleic acid at a concentration of about 2.13 mg/kg, and a sterol at a concentration of about 1.01 mg/kg (wherein mg/kg=parts per million). In certain embodiments, the T-cell expansion composition comprises octanoic acid at a concentration of 9.19 mg/kg, palmitic acid at a concentration of 1.86 mg/kg, linoleic acid at a concentration of 2.12 mg/kg, oleic acid at a concentration of about 2.13 mg/kg, and a sterol at a concentration of 1.01 mg/kg (wherein mg/kg=parts per million). In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of between 6.4 μmol/kg and 640 μmol/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.7 μmol/kg and 70 μmol/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.75 μmol/kg and 75 μmol/kg, inclusive of the endpoints; oleic acid at a concentration of between 0.75 μmol/kg and 75 μmol/kg, inclusive of the endpoints; and a sterol at a concentration of between 0.25 μmol/kg and 25 μmol/kg, inclusive of the endpoints. In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of about 64 μmol/kg, palmitic acid at a concentration of about 7 μmol/kg, linoleic acid at a concentration of about 7.5 μmol/kg, oleic acid at a concentration of about 7.5 μmol/kg and a sterol at a concentration of about 2.5 μmol/kg. In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of about 63.75 μmol/kg, palmitic acid at a concentration of about 7.27 μmol/kg, linoleic acid at a concentration of about 7.57 μmol/kg, oleic acid at a concentration of about 7.56 μmol/kg and a sterol at a concentration of about 2.61 μmol/kg. In certain embodiments, the T-cell expansion composition comprises octanoic acid at a concentration of about 63.75 μmol/kg, palmitic acid at a concentration of about 7.27 μmol/kg, linoleic acid at a concentration of about 7.57 μmol/kg, oleic acid at a concentration of 7.56 μmol/kg and a sterol at a concentration of 2.61 μmol/kg.

As used herein, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, and an expansion supplement at 37° C. Alternatively, or in addition, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of the following components: octanoic acid (CAS No. 124-07-2), nicotinamide (CAS No. 98-92-0), 2,4,7,9-tetramethyl-5-decyn-4,7-diol (TMDD) (CAS No. 126-86-3), diisopropyl adipate (DIPA) (CAS No. 6938-94-9), n-butyl-benzenesulfonamide (CAS No. 3622-84-2), 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester (CAS No. 84-69-5), palmitic acid (CAS No. 57-10-3), linoleic acid (CAS No. 60-33-3), oleic acid (CAS No. 112-80-1), stearic acid hydrazide (CAS No. 4130-54-5), oleamide (CAS No. 3322-62-1), sterol (e.g., cholesterol) (CAS No. 57-88-5), and alkanes (e.g., nonadecane) (CAS No. 629-92-5). In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of the following components: octanoic acid (CAS No. 124-07-2), nicotinamide (CAS No. 98-92-0), 2,4,7,9-tetramethyl-5-decyn-4,7-diol (TMDD) (CAS No. 126-86-3), diisopropyl adipate (DIPA) (CAS No. 6938-94-9), n-butyl-benzenesulfonamide (CAS No. 3622-84-2), 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester (CAS No. 84-69-5), palmitic acid (CAS No. 57-10-3), linoleic acid (CAS No. 60-33-3), oleic acid (CAS No. 112-80-1), stearic acid hydrazide (CAS No. 4130-54-5), oleamide (CAS No. 3322-62-1), sterol (e.g., cholesterol) (CAS No. 57-88-5), alkanes (e.g., nonadecane) (CAS No. 629-92-5), and phenol red (CAS No. 143-74-8). In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of the following components: octanoic acid (CAS No. 124-07-2), nicotinamide (CAS No. 98-92-0), 2,4,7,9-tetramethyl-5-decyn-4,7-diol (TMDD) (CAS No. 126-86-3), diisopropyl adipate (DIPA) (CAS No. 6938-94-9), n-butyl-benzenesulfonamide (CAS No. 3622-84-2), 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester (CAS No. 84-69-5), palmitic acid (CAS No. 57-10-3), linoleic acid (CAS No. 60-33-3), oleic acid (CAS No. 112-80-1), stearic acid hydrazide (CAS No. 4130-54-5), oleamide (CAS No. 3322-62-1), phenol red (CAS No. 143-74-8) and lanolin alcohol.

In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, and an expansion supplement at 37° C. Alternatively, or in addition, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of the following ions: sodium, ammonium, potassium, magnesium, calcium, chloride, sulfate and phosphate.

As used herein, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, and an expansion supplement at 37° C. Alternatively, or in addition, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of the following free amino acids: histidine, asparagine, serine, glutamate, arginine, glycine, aspartic acid, glutamic acid, threonine, alanine, proline, cysteine, lysine, tyrosine, methionine, valine, isoleucine, leucine, phenylalanine and tryptophan. In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of the following free amino acids in the corresponding average mole percentages: histidine (about 1%), asparagine (about 0.5%), serine (about 1.5%), glutamine (about 67%), arginine (about 1.5%), glycine (about 1.5%), aspartic acid (about 1%), glutamic acid (about 2%), threonine (about 2%), alanine (about 1%), proline (about 1.5%), cysteine (about 1.5%), lysine (about 3%), tyrosine (about 1.5%), methionine (about 1%), valine (about 3.5%), isoleucine (about 3%), leucine (about 3.5%), phenylalanine (about 1.5%) and tryptophan (about 0.5%). In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of the following free amino acids in the corresponding average mole percentages: histidine (about 0.78%), asparagine (about 0.4%), serine (about 1.6%), glutamine (about 67.01%), arginine (about 1.67%), glycine (about 1.72%), aspartic acid (about 1.00%), glutamic acid (about 1.93%), threonine (about 2.38%), alanine (about 1.11%), proline (about 1.49%), cysteine (about 1.65%), lysine (about 2.84%), tyrosine (about 1.62%), methionine (about 0.85%), valine (about 3.45%), isoleucine (about 3.14%), leucine (about 3.3%), phenylalanine (about 1.64%) and tryptophan (about 0.37%).

In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of octanoic acid, palmitic acid, linoleic acid, oleic acid and a sterol (e.g. cholesterol). In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of between 0.9 mg/kg to 90 mg/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; oleic acid at a concentration of 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; and a sterol at a concentration of about 0.1 mg/kg to 10 mg/kg, inclusive of the endpoints (wherein mg/kg=parts per million). In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of about 9 mg/kg, palmitic acid at a concentration of about 2 mg/kg, linoleic acid at a concentration of about 2 mg/kg, oleic acid at a concentration of about 2 mg/kg, and a sterol at a concentration of about 1 mg/kg (wherein mg/kg=parts per million). In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of 9.19 mg/kg, palmitic acid at a concentration of 1.86 mg/kg, linoleic acid at a concentration of about 2.12 mg/kg, oleic acid at a concentration of about 2.13 mg/kg, and a sterol at a concentration of about 1.01 mg/kg (wherein mg/kg=parts per million). In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of 9.19 mg/kg, palmitic acid at a concentration of 1.86 mg/kg, linoleic acid at a concentration of 2.12 mg/kg, oleic acid at a concentration of about 2.13 mg/kg, and a sterol at a concentration of 1.01 mg/kg (wherein mg/kg=parts per million). In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of between 6.4 μmol/kg and 640 μmol/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.7 μmol/kg and 70 μmol/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.75 μmol/kg and 75 μmol/kg, inclusive of the endpoints; oleic acid at a concentration of between 0.75 μmol/kg and 75 μmol/kg, inclusive of the endpoints; and a sterol at a concentration of between 0.25 μmol/kg and 25 μmol/kg, inclusive of the endpoints. In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of about 64 μmol/kg, palmitic acid at a concentration of about 7 μmol/kg, linoleic acid at a concentration of about 7.5 μmol/kg, oleic acid at a concentration of about 7.5 μmol/kg and a sterol at a concentration of about 2.5 μmol/kg.

In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of about 63.75 μmol/kg, palmitic acid at a concentration of about 7.27 μmol/kg, linoleic acid at a concentration of about 7.57 μmol/kg, oleic acid at a concentration of about 7.56 μmol/kg and a sterol at a concentration of about 2.61 μmol/kg. In certain embodiments, the terms “supplemented T-cell expansion composition” or “T-cell expansion composition” may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of about 63.75 μmol/kg, palmitic acid at a concentration of about 7.27 μmol/kg, linoleic acid at a concentration of about 7.57 μmol/kg, oleic acid at a concentration of 7.56 μmol/kg and a sterol at a concentration of 2.61 μmol/kg.

Modified T-cells of the disclosure, including modified stem cell-like T cells, T_SCMand/or T_CMof the disclosure, may be incubated, cultured, grown, stored, or otherwise, combined at any step in the methods of the procedure with a growth medium comprising one or more inhibitors a component of a PI3K pathway. Exemplary inhibitors a component of a PI3K pathway include, but are not limited to, an inhibitor of GSK3β such as TWS119 (also known as GSK 3B inhibitor XII; CAS Number 601514-19-6 having a chemical formula C₁₈H₁₄N₄O₂). Exemplary inhibitors of a component of a PI3K pathway include, but are not limited to, bb007 (BLUEBIRDBIO™).

In some embodiments of the methods of the disclosure, the methods comprise contacting an immune cell of the disclosure and a T-cell activator composition. In some embodiments of the methods of the disclosure, the methods comprise contacting an immune cell precursor of the disclosure and a T-cell activator composition. In some embodiments of the methods of the disclosure, the methods comprise contacting a modified T cell of the disclosure and a T-cell activator composition. In some embodiments, the T-cell activator composition comprises one or more of an anti-human CD3 monospecific tetrameric antibody complex, an anti-human CD28 monospecific tetrameric antibody complex and an activation supplement to produce an activated modified T-cell or a plurality of activated modified T-cells. In some embodiments, the activated modified T-cell expresses one or more cell-surface marker(s) of an early memory T cell, a stem cell-like T cell, a T_SCMor a T_CM. In some embodiments, at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of activated modified T-cells express one or more cell-surface marker(s) of an early memory T cell, a stem cell-like T cell, a T_SCMor a T_CM.

In certain embodiments of the methods of producing a modified T cell (e.g. a stem cell-like T cell, a T_SCMand/or a T_CM) of the disclosure, the activation supplement may comprise one or more cytokine(s). The one or more cytokine(s) may comprise any cytokine, including but not limited to, lymphokines. Exemplary lympokines include, but are not limited to, interleukin-2 (IL-2), interleukin-3 (IL-3), interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-6 (IL-6), interleukin-7 (IL-7), interleukin-15 (IL-15), interleukin-21 (IL-21), granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma (INFγ). The one or more cytokine(s) may comprise IL-2.

Natural Killer (NK) Cells

In certain embodiments, the modified immune or immune precursor cells of the disclosure are natural killer (NK) cells. In certain embodiments, NK cells are cytotoxic lymphocytes that differentiate from lymphoid progenitor cells.

Modified NK cells of the disclosure may be derived from modified hematopoietic stem and progenitor cells (HSPCs) or modified HSCs.

In certain embodiments, non-activated NK cells are derived from CD3-depleted leukopheresis (containing CD14/CD19/CD56+ cells).

In certain embodiments, NK cells are electroporated using a Lonza 4D nucleofector or BTX ECM 830 (500V, 700 usec pulse length, 0.2 mm electrode gap, one pulse). All Lonza 4D nucleofector programs are contemplated as within the scope of the methods of the disclosure.

In certain embodiments, 5×10E6 cells were electroporated per electroporation in 100 μL P3 buffer in cuvettes. However, this ratio of cells per volume is scalable for commercial manufacturing methods.

In certain embodiments, NK cells were stimulated by co-culture with an additional cell line. In certain embodiments, the additional cell line comprises artificial antigen presenting cells (aAPCs). In certain embodiments, stimulation occurs at day 1, 2, 3, 4, 5, 6, or 7 following electroporation. In certain embodiments, stimulation occurs at day 2 following electroporation.

In certain embodiments, NK cells express CD56.

B cells

In certain embodiments, the modified immune or immune precursor cells of the disclosure are B cells. B cells are a type of lymphocyte that express B cell receptors on the cell surface. B cell receptors bind to specific antigens.

Modified B cells of the disclosure may be derived from modified hematopoietic stem and progenitor cells (HSPCs) or modified HSCs.

In certain embodiments, HSPCs are modified using the methods of the disclosure, and then primed for B cell differentiation in presence of human IL-3, Flt3L, TPO, SCF, and G-CSF for at least 3 days, at least 4 days, at least 5 days, at least 6 days or at least 7 days. In certain embodiments, HSPCs are modified using the methods of the disclosure, and then primed for B cell differentiation in presence of human IL-3, Flt3L, TPO, SCF, and G-CSF for 5 days.

In certain embodiments, following priming, modified HSPC cells are transferred to a layer of feeder cells and fed bi-weekly, along with transfer to a fresh layer of feeders once per week. In certain embodiments, the feeder cells are MS-5 feeder cells.

In certain embodiments, modified HSPC cells are cultured with MS-5 feeder cells for at least 7, 14, 21, 28, 30, 33, 35, 42 or 48 days. In certain embodiments, modified HSPC cells were cultured with MS-5 feeder cells for 33 days.

Chimeric Antigen Receptors

In certain embodiments, a modified immune or pre-immune cell of the disclosure comprises a chimeric antigen receptor.

In certain embodiments of the methods of the disclosure, the recombinant and non-naturally occurring DNA sequence encoding a transposon further comprises a sequence encoding a chimeric antigen receptor or a portion thereof. Chimeric antigen receptors (CARs) of the disclosure may comprise (a) an ectodomain comprising an antigen recognition region, (b) a transmembrane domain, and (c) an endodomain comprising at least one costimulatory domain. In certain embodiments, the ectodomain may further comprise a signal peptide. Alternatively, or in addition, in certain embodiments, the ectodomain may further comprise a hinge between the antigen recognition region and the transmembrane domain. In certain embodiments of the CARs of the disclosure, the signal peptide may comprise a sequence encoding a human CD2, CD3δ, CD3ε, CD3γ, CD3ζ, CD4, CD8α, CD19, CD28, 4-1BB or GM-CSFR signal peptide. In certain embodiments of the CARs of the disclosure, the signal peptide may comprise a sequence encoding a human CD8α signal peptide. In certain embodiments, the transmembrane domain may comprise a sequence encoding a human CD2, CD3δ, CD3ε, CD3γ, CD3ζ, CD4, CD8α, CD19, CD28, 4-1BB or GM-CSFR transmembrane domain. In certain embodiments of the CARs of the disclosure, the transmembrane domain may comprise a sequence encoding a human CD8α transmembrane domain. In certain embodiments of the CARs of the disclosure, the endodomain may comprise a human CD3 endodomain.

In certain embodiments of the CARs of the disclosure, the at least one costimulatory domain may comprise a human 4-1BB, CD28, CD40, ICOS, MyD88, OX-40 intracellular segment, or any combination thereof. In certain embodiments of the CARs of the disclosure, the at least one costimulatory domain may comprise a CD28 and/or a 4-1BB costimulatory domain. In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD8α, IgG4, and/or CD4 sequence. In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD8α sequence.

The CD28 costimulatory domain may comprise an amino acid sequence comprising

(SEQ ID NO: 14659)

RVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPR

RKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDT

YDALHMQALPPR

or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising

(SEQ ID NO: 14659)

RVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPR

RKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDT

YDALHMQALPPR.

The CD28 costimulatory domain may be encoded by the nucleic acid sequence comprising

(SEQ ID NO: 14660)

cgcgtgaagtttagtcgatcagcagatgccccagcttacaaacagggaca

gaaccagctgtataacgagctgaatctgggccgccgagaggaatatgacg

tgctggataagcggagaggacgcgaccccgaaatgggaggcaagcccagg

cgcaaaaaccctcaggaaggcctgtataacgagctgcagaaggacaaaat

ggcagaagcctattctgagatcggcatgaagggggagcgacggagaggca

aagggcacgatgggctgtaccagggactgagcaccgccacaaaggacacc

tatgatgctctgcatatgcaggcactgcctccaagg.

The 4-1BB costimulatory domain may comprise an amino acid sequence comprising

(SEQ ID NO: 14661)

KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL

or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising

(SEQ ID NO: 14661)

KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL.

The 4-1BB costimulatory domain may be encoded by the nucleic acid sequence comprising

(SEQ ID NO: 14662)

aagagaggcaggaagaaactgctgtatattttcaaacagcccttcatgcg

ccccgtgcagactacccaggaggaagacgggtgctcctgtcgattccctg

aggaagaggaaggcgggtgtgagctg.

The 4-1BB costimulatory domain may be located between the transmembrane domain and the CD28 costimulatory domain.

In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD8α, IgG4, and/or CD4 sequence. In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD8α sequence. The hinge may comprise a human CD8α amino acid sequence comprising

(SEQ ID NO: 14663)

TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD

or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising

(SEQ ID NO: 14663)

TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD.

The human CD8α hinge amino acid sequence may be encoded by the nucleic acid sequence comprising

(SEQ ID NO: 14664)

actaccacaccagcacctagaccaccaactccagctccaaccatcgcgag

tcagcccctgagtctgagacctgaggcctgcaggccagctgcaggaggag

ctgtgcacaccaggggcctggacttcgcctgcgac.

ScFv

The disclosure provides single chain variable fragment (scFv) compositions and methods for use of these compositions to recognize and bind to a specific target protein. ScFv compositions comprise a heavy chain variable region and a light chain variable region of an antibody. ScFv compositions may be incorporated into an antigen recognition region of a chimeric antigen receptor of the disclosure. ScFvs are fusion proteins of the variable regions of the heavy (VH) and light (VL) chains of immunoglobulins, and the VH and VL domains are connected with a short peptide linker. ScFvs retain the specificity of the original immunoglobulin, despite removal of the constant regions and the introduction of the linker. An exemplary linker comprises a sequence of GGGGSGGGGSGGGGS (SEQ ID NO: 14665).

Centyrins

Centyrins of the disclosure specifically bind to an antigen. Chimeric antigen receptors of the disclosure comprising one or more Centyrins that specifically bind an antigen may be used to direct the specificity of a cell, (e.g. a cytotoxic immune cell) towards the specific antigen.

Centyrins of the disclosure may comprise a protein scaffold, wherein the scaffold is capable of specifically binding an antigen. Centyrins of the disclosure may comprise a protein scaffold comprising a consensus sequence of at least one fibronectin type III (FN3) domain, wherein the scaffold is capable of specifically binding an antigen. The at least one fibronectin type III (FN3) domain may be derived from a human protein. The human protein may be Tenascin-C. The consensus sequence may comprise

(SEQ ID NO: 14488)

LPAPKNLVVSEVTEDSLRLSWTAPDAAFDSFLIQYQESEKVGEAINLTVP

GSERSYDLTGLKPGTEYTVSIYGVKGGHRSNPLSAEFTT

or

MLPAPKNLVVSEVTEDSLRLSWTAPDAAFDSFLIQYQESEKVGEAINLTV

PGSERSYD

The consensus sequence may comprise an amino sequence at least 74% identical to

(SEQ ID NO: 14488)

LPAPKNLVVSEVTEDSLRLSWTAPDAAFDSFLIQYQESEKVGEAINLTVP

GSERSYDLTGLKPGTEYTVSIYGVKGGHRSNPLSAEFTT

or

(SEQ ID NO: 14489)

MLPAPKNLVVSEVTEDSLRLSWTAPDAAFDSFLIQYQESEKVGEAINLTV

PGSERSYDLTGLKPGTEYTVSIYGVKGGHRSNPLSAEFTT.

The consensus sequence may encoded by a nucleic acid sequence comprising

(SEQ ID NO: 14490)

atgctgcctgcaccaaagaacctggtggtgtctcatgtgacagaggatag

tgccagactgtcatggactgctcccgacgcagccttcgatagttttatca

tcgtgtaccgggagaacatcgaaaccggcgaggccattgtcctgacagtg

ccagggtccgaacgctcttatgacctgacagatctgaagcccggaactga

gtactatgtgcagatcgccggcgtcaaaggaggcaatatcagcttccctc

tgtccgcaatcttcaccaca.

The consensus sequence may be modified at one or more positions within (a) a A-B loop comprising or consisting of the amino acid residues TEDS (SEQ ID NO: 14491) at positions 13-16 of the consensus sequence; (b) a B-C loop comprising or consisting of the amino acid residues TAPDAAF (SEQ ID NO: 14492) at positions 22-28 of the consensus sequence; (c) a C-D loop comprising or consisting of the amino acid residues SEKVGE (SEQ ID NO: 14493) at positions 38-43 of the consensus sequence; (d) a D-E loop comprising or consisting of the amino acid residues GSER (SEQ ID NO: 14494) at positions 51-54 of the consensus sequence; (e) a E-F loop comprising or consisting of the amino acid residues GLKPG (SEQ ID NO: 14495) at positions 60-64 of the consensus sequence; (f) a F-G loop comprising or consisting of the amino acid residues KGGHRSN (SEQ ID NO: 14496) at positions 75-81 of the consensus sequence; or (g) any combination of (a)-(f). Centyrins of the disclosure may comprise a consensus sequence of at least 5 fibronectin type III (FN3) domains, at least 10 fibronectin type III (FN3) domains or at least 15 fibronectin type III (FN3) domains. The scaffold may bind an antigen with at least one affinity selected from a K_Dof less than or equal to 10M, less than or equal to 10⁻¹⁰M, less than or equal to 10⁻¹¹M, less than or equal to 10⁻¹²M, less than or equal to 10⁻¹³M, less than or equal to 10⁻¹⁴M, and less than or equal to 10⁻¹⁵M. The K_Dmay be determined by surface plasmon resonance.

The term “antibody mimetic” is intended to describe an organic compound that specifically binds a target sequence and has a structure distinct from a naturally-occurring antibody. Antibody mimetics may comprise a protein, a nucleic acid, or a small molecule. The target sequence to which an antibody mimetic of the disclosure specifically binds may be an antigen. Antibody mimetics may provide superior properties over antibodies including, but not limited to, superior solubility, tissue penetration, stability towards heat and enzymes (e.g. resistance to enzymatic degradation), and lower production costs. Exemplary antibody mimetics include, but are not limited to, an affibody, an afflilin, an affimer, an affitin, an alphabody, an anticalin, and avimer (also known as avidity multimer), a DARPin (Designed Ankyrin Repeat Protein), a Fynomer, a Kunitz domain peptide, and a monobody.

Affibody molecules of the disclosure comprise a protein scaffold comprising or consisting of one or more alpha helix without any disulfide bridges. Preferably, affibody molecules of the disclosure comprise or consist of three alpha helices. For example, an affibody molecule of the disclosure may comprise an immunoglobulin binding domain. An affibody molecule of the disclosure may comprise the Z domain of protein A.

Affilin molecules of the disclosure comprise a protein scaffold produced by modification of exposed amino acids of, for example, either gamma-B crystallin or ubiquitin. Affilin molecules functionally mimic an antibody's affinity to antigen, but do not structurally mimic an antibody. In any protein scaffold used to make an affilin, those amino acids that are accessible to solvent or possible binding partners in a properly-folded protein molecule are considered exposed amino acids. Any one or more of these exposed amino acids may be modified to specifically bind to a target sequence or antigen.

Affimer molecules of the disclosure comprise a protein scaffold comprising a highly stable protein engineered to display peptide loops that provide a high affinity binding site for a specific target sequence. Exemplary affimer molecules of the disclosure comprise a protein scaffold based upon a cystatin protein or tertiary structure thereof. Exemplary affimer molecules of the disclosure may share a common tertiary structure of comprising an alpha-helix lying on top of an anti-parallel beta-sheet.

Affitin molecules of the disclosure comprise an artificial protein scaffold, the structure of which may be derived, for example, from a DNA binding protein (e.g. the DNA binding protein Sac7d). Affitins of the disclosure selectively bind a target sequence, which may be the entirety or part of an antigen. Exemplary affitins of the disclosure are manufactured by randomizing one or more amino acid sequences on the binding surface of a DNA binding protein and subjecting the resultant protein to ribosome display and selection. Target sequences of affitins of the disclosure may be found, for example, in the genome or on the surface of a peptide, protein, virus, or bacteria. In certain embodiments of the disclosure, an affitin molecule may be used as a specific inhibitor of an enzyme. Affitin molecules of the disclosure may include heat-resistant proteins or derivatives thereof.

Alphabody molecules of the disclosure may also be referred to as Cell-Penetrating Alphabodies (CPAB). Alphabody molecules of the disclosure comprise small proteins (typically of less than 10 kDa) that bind to a variety of target sequences (including antigens). Alphabody molecules are capable of reaching and binding to intracellular target sequences. Structurally, alphabody molecules of the disclosure comprise an artificial sequence forming single chain alpha helix (similar to naturally occurring coiled-coil structures). Alphabody molecules of the disclosure may comprise a protein scaffold comprising one or more amino acids that are modified to specifically bind target proteins. Regardless of the binding specificity of the molecule, alphabody molecules of the disclosure maintain correct folding and thermostability.

Anticalin molecules of the disclosure comprise artificial proteins that bind to target sequences or sites in either proteins or small molecules. Anticalin molecules of the disclosure may comprise an artificial protein derived from a human lipocalin. Anticalin molecules of the disclosure may be used in place of, for example, monoclonal antibodies or fragments thereof. Anticalin molecules may demonstrate superior tissue penetration and thermostability than monoclonal antibodies or fragments thereof. Exemplary anticalin molecules of the disclosure may comprise about 180 amino acids, having a mass of approximately 20 kDa. Structurally, anticalin molecules of the disclosure comprise a barrel structure comprising antiparallel beta-strands pairwise connected by loops and an attached alpha helix. In preferred embodiments, anticalin molecules of the disclosure comprise a barrel structure comprising eight antiparallel beta-strands pairwise connected by loops and an attached alpha helix.

Avimer molecules of the disclosure comprise an artificial protein that specifically binds to a target sequence (which may also be an antigen). Avimers of the disclosure may recognize multiple binding sites within the same target or within distinct targets. When an avimer of the disclosure recognize more than one target, the avimer mimics function of a bi-specific antibody. The artificial protein avimer may comprise two or more peptide sequences of approximately 30-35 amino acids each. These peptides may be connected via one or more linker peptides. Amino acid sequences of one or more of the peptides of the avimer may be derived from an A domain of a membrane receptor. Avimers have a rigid structure that may optionally comprise disulfide bonds and/or calcium. Avimers of the disclosure may demonstrate greater heat stability compared to an antibody.

DARPins (Designed Ankyrin Repeat Proteins) of the disclosure comprise genetically-engineered, recombinant, or chimeric proteins having high specificity and high affinity for a target sequence. In certain embodiments, DARPins of the disclosure are derived from ankyrin proteins and, optionally, comprise at least three repeat motifs (also referred to as repetitive structural units) of the ankyrin protein. Ankyrin proteins mediate high-affinity protein-protein interactions. DARPins of the disclosure comprise a large target interaction surface.

Fynomers of the disclosure comprise small binding proteins (about 7 kDa) derived from the human Fyn SH3 domain and engineered to bind to target sequences and molecules with equal affinity and equal specificity as an antibody.

Kunitz domain peptides of the disclosure comprise a protein scaffold comprising a Kunitz domain. Kunitz domains comprise an active site for inhibiting protease activity. Structurally, Kunitz domains of the disclosure comprise a disulfide-rich alpha+beta fold. This structure is exemplified by the bovine pancreatic trypsin inhibitor. Kunitz domain peptides recognize specific protein structures and serve as competitive protease inhibitors. Kunitz domains of the disclosure may comprise Ecallantide (derived from a human lipoprotein-associated coagulation inhibitor (LACI)).

Monobodies of the disclosure are small proteins (comprising about 94 amino acids and having a mass of about 10 kDa) comparable in size to a single chain antibody. These genetically engineered proteins specifically bind target sequences including antigens. Monobodies of the disclosure may specifically target one or more distinct proteins or target sequences. In preferred embodiments, monobodies of the disclosure comprise a protein scaffold mimicking the structure of human fibronectin, and more preferably, mimicking the structure of the tenth extracellular type III domain of fibronectin. The tenth extracellular type III domain of fibronectin, as well as a monobody mimetic thereof, contains seven beta sheets forming a barrel and three exposed loops on each side corresponding to the three complementarity determining regions (CDRs) of an antibody. In contrast to the structure of the variable domain of an antibody, a monobody lacks any binding site for metal ions as well as a central disulfide bond. Multispecific monobodies may be optimized by modifying the loops BC and FG. Monobodies of the disclosure may comprise an adnectin.

VHH

In certain embodiments, the CAR comprises a single domain antibody (SdAb). In certain embodiments, the SdAb is a VHH.

The disclosure provides chimeric antigen receptors (CARs) comprising at least one VHH (a VCAR). Chimeric antigen receptors of the disclosure may comprise more than one VHH. For example, a bi-specific VCAR may comprise two VHHs that specifically bind two distinct antigens.

VHH proteins of the disclosure specifically bind to an antigen. Chimeric antigen receptors of the disclosure comprising one or more VHHs that specifically bind an antigen may be used to direct the specificity of a cell, (e.g. a cytotoxic immune cell) towards the specific antigen.

At least one VHH protein or VCAR of the disclosure can be optionally produced by a cell line, a mixed cell line, an immortalized cell or clonal population of immortalized cells, as well known in the art. See, e.g., Ausubel, et al., ed., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., NY, N.Y. (1987-2001); Sambrook, et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor, N.Y. (1989); Harlow and Lane, Antibodies, a Laboratory Manual, Cold Spring Harbor, N.Y. (1989); Colligan, et al., eds., Current Protocols in Immunology, John Wiley & Sons, Inc., NY (1994-2001); Colligan et al., Current Protocols in Protein Science, John Wiley & Sons, NY, N.Y., (1997-2001).

Amino acids from a VHH protein can be altered, added and/or deleted to reduce immunogenicity or reduce, enhance or modify binding, affinity, on-rate, off-rate, avidity, specificity, half-life, stability, solubility or any other suitable characteristic, as known in the art.

Optionally, VHH proteins can be engineered with retention of high affinity for the antigen and other favorable biological properties. To achieve this goal, the VHH proteins can be optionally prepared by a process of analysis of the parental sequences and various conceptual engineered products using three-dimensional models of the parental and engineered sequences. Three-dimensional models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate sequences and can measure possible immunogenicity (e.g., Immunofilter program of Xencor, Inc. of Monrovia, Calif.). Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate sequence, i.e., the analysis of residues that influence the ability of the candidate VHH protein to bind its antigen. In this way, residues can be selected and combined from the parent and reference sequences so that the desired characteristic, such as affinity for the target antigen(s), is achieved. Alternatively, or in addition to, the above procedures, other suitable methods of engineering can be used.

Screening VHH for specific binding to similar proteins or fragments can be conveniently achieved using nucleotide (DNA or RNA display) or peptide display libraries, for example, in vitro display. This method involves the screening of large collections of peptides for individual members having the desired function or structure. The displayed nucleotide or peptide sequences can be from 3 to 5000 or more nucleotides or amino acids in length, frequently from 5-100 amino acids long, and often from about 8 to 25 amino acids long. In addition to direct chemical synthetic methods for generating peptide libraries, several recombinant DNA methods have been described. One type involves the display of a peptide sequence on the surface of a bacteriophage or cell. Each bacteriophage or cell contains the nucleotide sequence encoding the particular displayed peptide sequence. The VHH proteins of the disclosure can bind human or other mammalian proteins with a wide range of affinities (KD). In a preferred embodiment, at least one VHH of the present invention can optionally bind to a target protein with high affinity, for example, with a KD equal to or less than about 10⁻⁷M, such as but not limited to, 0.1-9.9 (or any range or value therein)×10⁻⁸, 10⁻⁹, 10⁻¹⁰, 10⁻¹¹, 10⁻¹², 10⁻¹³, 10⁻¹⁴, 10⁻¹⁵or any range or value therein, as determined by surface plasmon resonance or the Kinexa method, as practiced by those of skill in the art.

The affinity or avidity of a VHH or a VCAR for an antigen can be determined experimentally using any suitable method. (See, for example, Berzofsky, et al., “Antibody-Antigen Interactions,” In Fundamental Immunology, Paul, W. E., Ed., Raven Press: New York, N.Y. (1984); Kuby, Janis Immunology, W.H. Freeman and Company: New York, N.Y. (1992); and methods described herein). The measured affinity of a particular VHH-antigen or VCAR-antigen interaction can vary if measured under different conditions (e.g., salt concentration, pH). Thus, measurements of affinity and other antigen-binding parameters (e.g., KD, Kon, Koff) are preferably made with standardized solutions of VHH or VCAR and antigen, and a standardized buffer, such as the buffer described herein.

Competitive assays can be performed with the VHH or VCAR of the disclosure in order to determine what proteins, antibodies, and other antagonists compete for binding to a target protein with the VHH or VCAR of the present invention and/or share the epitope region. These assays as readily known to those of ordinary skill in the art evaluate competition between antagonists or ligands for a limited number of binding sites on a protein. The protein and/or antibody is immobilized or insolubilized before or after the competition and the sample bound to the target protein is separated from the unbound sample, for example, by decanting (where the protein/antibody was preinsolubilized) or by centrifuging (where the protein/antibody was precipitated after the competitive reaction). Also, the competitive binding may be determined by whether function is altered by the binding or lack of binding of the VHH or VCAR to the target protein, e.g., whether the VCAR molecule inhibits or potentiates the enzymatic activity of, for example, a label. ELISA and other functional assays may be used, as well known in the art.

In certain embodiments, the CAR comprises a single domain antibody (SdAb). In certain embodiments, the SdAb is a VH.

The disclosure provides chimeric antigen receptors (CARs) comprising a single domain antibody (VCARs). In certain embodiments, the single domain antibody comprises a VH. In certain embodiments, the VH is isolated or derived from a human sequence. In certain embodiments, VH comprises a human CDR sequence and/or a human framework sequence and a non-human or humanized sequence (e.g. a rat Fc domain). In certain embodiments, the VH is a fully humanized VH. In certain embodiments, the VH s neither a naturally occurring antibody nor a fragment of a naturally occurring antibody. In certain embodiments, the VH is not a fragment of a monoclonal antibody. In certain embodiments, the VH is a UniDab™ antibody (TeneoBio).

In certain embodiments, the VH is fully engineered using the UniRat™ (TeneoBio) system and “NGS-based Discovery” to produce the VH. Using this method, the specific VH are not naturally-occurring and are generated using fully engineered systems. The VH are not derived from naturally-occurring monoclonal antibodies (mAbs) that were either isolated directly from the host (for example, a mouse, rat or human) or directly from a single clone of cells or cell line (hybridoma). These VHs were not subsequently cloned from said cell lines. Instead, VH sequences are fully-engineered using the UniRat™ system as transgenes that comprise human variable regions (VH domains) with a rat Fc domain, and are thus human/rat chimeras without a light chain and are unlike the standard mAb format. The native rat genes are knocked out and the only antibodies expressed in the rat are from transgenes with VH domains linked to a Rat Fc (UniAbs). These are the exclusive Abs expressed in the UniRat. Next generation sequencing (NGS) and bioinformatics are used to identify the full antigen-specific repertoire of the heavy-chain antibodies generated by UniRat™ after immunization. Then, a unique gene assembly method is used to convert the antibody repertoire sequence information into large collections of fully-human heavy-chain antibodies that can be screened in vitro for a variety of functions. In certain embodiments, fully humanized VH are generated by fusing the human VH domains with human Fcs in vitro (to generate a non-naturally occurring recombinant VH antibody). In certain embodiments, the VH are fully humanized, but they are expressed in vivo as human/rat chimera (human VH, rat Fc) without a light chain. Fully humanized VHs are expressed in vivo as human/rat chimera (human VH, rat Fc) without a light chain are about 80 kDa (vs 150 kDa).

VCARs of the disclosure may comprise at least one VH of the disclosure. In certain embodiments, the VH of the disclosure may be modified to remove an Fc domain or a portion thereof. In certain embodiments, a framework sequence of the VH of the disclosure may be modified to, for example, improve expression, decrease immunogenicity or to improve function.

As used throughout the disclosure, the singular forms “a,” “and,” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a method” includes a plurality of such methods and reference to “a dose” includes reference to one or more doses and equivalents thereof known to those skilled in the art, and so forth.

The term “about” or “approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, “about” can mean within 1 or more standard deviations. Alternatively, “about” can mean a range of up to 20%, or up to 10%, or up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold, of a value. Where particular values are described in the application and claims, unless otherwise stated the term “about” meaning within an acceptable error range for the particular value should be assumed.

The disclosure provides isolated or substantially purified polynucleotide or protein compositions. An “isolated” or “purified” polynucleotide or protein, or biologically active portion thereof, is substantially or essentially free from components that normally accompany or interact with the polynucleotide or protein as found in its naturally occurring environment. Thus, an isolated or purified polynucleotide or protein is substantially free of other cellular material or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized. Optimally, an “isolated” polynucleotide is free of sequences (optimally protein encoding sequences) that naturally flank the polynucleotide (i.e., sequences located at the 5′ and 3′ ends of the polynucleotide) in the genomic DNA of the organism from which the polynucleotide is derived. For example, in various embodiments, the isolated polynucleotide can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb, or 0.1 kb of nucleotide sequence that naturally flank the polynucleotide in genomic DNA of the cell from which the polynucleotide is derived. A protein that is substantially free of cellular material includes preparations of protein having less than about 30%, 20%, 10%, 5%, or 1% (by dry weight) of contaminating protein. When the protein of the invention or biologically active portion thereof is recombinantly produced, optimally culture medium represents less than about 30%, 20%, 10%, 5%, or 1% (by dry weight) of chemical precursors or non-protein-of-interest chemicals.

The disclosure provides fragments and variants of the disclosed DNA sequences and proteins encoded by these DNA sequences. As used throughout the disclosure, the term “fragment” refers to a portion of the DNA sequence or a portion of the amino acid sequence and hence protein encoded thereby. Fragments of a DNA sequence comprising coding sequences may encode protein fragments that retain biological activity of the native protein and hence DNA recognition or binding activity to a target DNA sequence as herein described. Alternatively, fragments of a DNA sequence that are useful as hybridization probes generally do not encode proteins that retain biological activity or do not retain promoter activity. Thus, fragments of a DNA sequence may range from at least about 20 nucleotides, about 50 nucleotides, about 100 nucleotides, and up to the full-length polynucleotide of the invention.

Nucleic acids or proteins of the disclosure can be constructed by a modular approach including preassembling monomer units and/or repeat units in target vectors that can subsequently be assembled into a final destination vector. Polypeptides of the disclosure may comprise repeat monomers of the disclosure and can be constructed by a modular approach by preassembling repeat units in target vectors that can subsequently be assembled into a final destination vector. The disclosure provides polypeptide produced by this method as well nucleic acid sequences encoding these polypeptides. The disclosure provides host organisms and cells comprising nucleic acid sequences encoding polypeptides produced this modular approach.

The term “antibody” is used in the broadest sense and specifically covers single monoclonal antibodies (including agonist and antagonist antibodies) and antibody compositions with polyepitopic specificity. It is also within the scope hereof to use natural or synthetic analogs, mutants, variants, alleles, homologs and orthologs (herein collectively referred to as “analogs”) of the antibodies hereof as defined herein. Thus, according to one embodiment hereof, the term “antibody hereof” in its broadest sense also covers such analogs. Generally, in such analogs, one or more amino acid residues may have been replaced, deleted and/or added, compared to the antibodies hereof as defined herein.

“Antibody fragment”, and all grammatical variants thereof, as used herein are defined as a portion of an intact antibody comprising the antigen binding site or variable region of the intact antibody, wherein the portion is free of the constant heavy chain domains (i.e. CH2, CH3, and CH4, depending on antibody isotype) of the Fc region of the intact antibody. Examples of antibody fragments include Fab, Fab′, Fab′-SH, F(ab′)₂, and Fv fragments; diabodies; any antibody fragment that is a polypeptide having a primary structure consisting of one uninterrupted sequence of contiguous amino acid residues (referred to herein as a “single-chain antibody fragment” or “single chain polypeptide”), including without limitation (l) single-chain Fv (scFv) molecules (2) single chain polypeptides containing only one light chain variable domain, or a fragment thereof that contains the three CDRs of the light chain variable domain, without an associated heavy chain moiety and (3) single chain polypeptides containing only one heavy chain variable region, or a fragment thereof containing the three CDRs of the heavy chain variable region, without an associated light chain moiety; and multispecific or multivalent structures formed from antibody fragments. In an antibody fragment comprising one or more heavy chains, the heavy chain(s) can contain any constant domain sequence (e.g. CHI in the IgG isotype) found in a non-Fc region of an intact antibody, and/or can contain any hinge region sequence found in an intact antibody, and/or can contain a leucine zipper sequence fused to or situated in the hinge region sequence or the constant domain sequence of the heavy chain(s). The term further includes single domain antibodies (“sdAB”) which generally refers to an antibody fragment having a single monomeric variable antibody domain, (for example, from camelids). Such antibody fragment types will be readily understood by a person having ordinary skill in the art.

“Binding” refers to a sequence-specific, non-covalent interaction between macromolecules (e.g., between a protein and a nucleic acid). Not all components of a binding interaction need be sequence-specific (e.g., contacts with phosphate residues in a DNA backbone), as long as the interaction as a whole is sequence-specific.

The term “comprising” is intended to mean that the compositions and methods include the recited elements, but do not exclude others. “Consisting essentially of” when used to define compositions and methods, shall mean excluding other elements of any essential significance to the combination when used for the intended purpose. Thus, a composition consisting essentially of the elements as defined herein would not exclude trace contaminants or inert carriers. “Consisting of shall mean excluding more than trace elements of other ingredients and substantial method steps. Embodiments defined by each of these transition terms are within the scope of this invention.

The term “epitope” refers to an antigenic determinant of a polypeptide. An epitope could comprise three amino acids in a spatial conformation, which is unique to the epitope. Generally, an epitope consists of at least 4, 5, 6, or 7 such amino acids, and more usually, consists of at least 8, 9, or 10 such amino acids. Methods of determining the spatial conformation of amino acids are known in the art, and include, for example, x-ray crystallography and two-dimensional nuclear magnetic resonance.

As used herein, “expression” refers to the process by which polynucleotides are transcribed into mRNA and/or the process by which the transcribed mRNA is subsequently being translated into peptides, polypeptides, or proteins. If the polynucleotide is derived from genomic DNA, expression may include splicing of the mRNA in a eukaryotic cell.

“Gene expression” refers to the conversion of the information, contained in a gene, into a gene product. A gene product can be the direct transcriptional product of a gene (e.g., mRNA, tRNA, rRNA, antisense RNA, ribozyme, shRNA, micro RNA, structural RNA or any other type of RNA) or a protein produced by translation of an mRNA. Gene products also include RNAs which are modified, by processes such as capping, polyadenylation, methylation, and editing, and proteins modified by, for example, methylation, acetylation, phosphorylation, ubiquitination, ADP-ribosylation, myristilation, and glycosylation.

“Modulation” or “regulation” of gene expression refers to a change in the activity of a gene. Modulation of expression can include, but is not limited to, gene activation and gene repression.

The term “operatively linked” or its equivalents (e.g., “linked operatively”) means two or more molecules are positioned with respect to each other such that they are capable of interacting to affect a function attributable to one or both molecules or a combination thereof.

Non-covalently linked components and methods of making and using non-covalently linked components, are disclosed. The various components may take a variety of different forms as described herein. For example, non-covalently linked (i.e., operatively linked) proteins may be used to allow temporary interactions that avoid one or more problems in the art. The ability of non-covalently linked components, such as proteins, to associate and dissociate enables a functional association only or primarily under circumstances where such association is needed for the desired activity. The linkage may be of duration sufficient to allow the desired effect.

A method for directing proteins to a specific locus in a genome of an organism is disclosed. The method may comprise the steps of providing a DNA localization component and providing an effector molecule, wherein the DNA localization component and the effector molecule are capable of operatively linking via a non-covalent linkage.

The term “scFv” refers to a single-chain variable fragment. scFv is a fusion protein of the variable regions of the heavy (VH) and light chains (VL) of immunoglobulins, connected with a linker peptide. The linker peptide may be from about 5 to 40 amino acids or from about 10 to 30 amino acids or about 5, 10, 15, 20, 25, 30, 35, or 40 amino acids in length. Single-chain variable fragments lack the constant Fc region found in complete antibody molecules, and, thus, the common binding sites (e.g., Protein G) used to purify antibodies. The term further includes a scFv that is an intrabody, an antibody that is stable in the cytoplasm of the cell, and which may bind to an intracellular protein.

The term “single domain antibody” means an antibody fragment having a single monomeric variable antibody domain which is able to bind selectively to a specific antigen. A single-domain antibody generally is a peptide chain of about 110 amino acids long, comprising one variable domain (VH) of a heavy-chain antibody, or of a common IgG, which generally have similar affinity to antigens as whole antibodies, but are more heat-resistant and stable towards detergents and high concentrations of urea. Examples are those derived from camelid or fish antibodies. Alternatively, single-domain antibodies can be made from common murine or human IgG with four chains.

The terms “specifically bind” and “specific binding” as used herein refer to the ability of an antibody, an antibody fragment or a nanobody to preferentially bind to a particular antigen that is present in a homogeneous mixture of different antigens. In certain embodiments, a specific binding interaction will discriminate between desirable and undesirable antigens in a sample. In certain embodiments more than about ten- to 100-fold or more (e.g., more than about 1000- or 10,000-fold). “Specificity” refers to the ability of an immunoglobulin or an immunoglobulin fragment, such as a nanobody, to bind preferentially to one antigenic target versus a different antigenic target and does not necessarily imply high affinity.

A “target site” or “target sequence” is a nucleic acid sequence that defines a portion of a nucleic acid to which a binding molecule will bind, provided sufficient conditions for binding exist.

The terms “nucleic acid” or “oligonucleotide” or “polynucleotide” refer to at least two nucleotides covalently linked together. The depiction of a single strand also defines the sequence of the complementary strand. Thus, a nucleic acid may also encompass the complementary strand of a depicted single strand. A nucleic acid of the disclosure also encompasses substantially identical nucleic acids and complements thereof that retain the same structure or encode for the same protein.

Probes of the disclosure may comprise a single stranded nucleic acid that can hybridize to a target sequence under stringent hybridization conditions. Thus, nucleic acids of the disclosure may refer to a probe that hybridizes under stringent hybridization conditions.

Nucleic acids of the disclosure may be single- or double-stranded. Nucleic acids of the disclosure may contain double-stranded sequences even when the majority of the molecule is single-stranded. Nucleic acids of the disclosure may contain single-stranded sequences even when the majority of the molecule is double-stranded. Nucleic acids of the disclosure may include genomic DNA, cDNA, RNA, or a hybrid thereof. Nucleic acids of the disclosure may contain combinations of deoxyribo- and ribo-nucleotides. Nucleic acids of the disclosure may contain combinations of bases including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine hypoxanthine, isocytosine and isoguanine. Nucleic acids of the disclosure may be synthesized to comprise non-natural amino acid modifications. Nucleic acids of the disclosure may be obtained by chemical synthesis methods or by recombinant methods.

Nucleic acids of the disclosure, either their entire sequence, or any portion thereof, may be non-naturally occurring. Nucleic acids of the disclosure may contain one or more mutations, substitutions, deletions, or insertions that do not naturally-occur, rendering the entire nucleic acid sequence non-naturally occurring. Nucleic acids of the disclosure may contain one or more duplicated, inverted or repeated sequences, the resultant sequence of which does not naturally-occur, rendering the entire nucleic acid sequence non-naturally occurring. Nucleic acids of the disclosure may contain modified, artificial, or synthetic nucleotides that do not naturally-occur, rendering the entire nucleic acid sequence non-naturally occurring.

Given the redundancy in the genetic code, a plurality of nucleotide sequences may encode any particular protein. All such nucleotides sequences are contemplated herein.

As used throughout the disclosure, the term “operably linked” refers to the expression of a gene that is under the control of a promoter with which it is spatially connected. A promoter can be positioned 5′ (upstream) or 3′ (downstream) of a gene under its control. The distance between a promoter and a gene can be approximately the same as the distance between that promoter and the gene it controls in the gene from which the promoter is derived. Variation in the distance between a promoter and a gene can be accommodated without loss of promoter function.

As used throughout the disclosure, the term “promoter” refers to a synthetic or naturally-derived molecule which is capable of conferring, activating or enhancing expression of a nucleic acid in a cell. A promoter can comprise one or more specific transcriptional regulatory sequences to further enhance expression and/or to alter the spatial expression and/or temporal expression of same. A promoter can also comprise distal enhancer or repressor elements, which can be located as much as several thousand base pairs from the start site of transcription. A promoter can be derived from sources including viral, bacterial, fungal, plants, insects, and animals. A promoter can regulate the expression of a gene component constitutively or differentially with respect to cell, the tissue or organ in which expression occurs or, with respect to the developmental stage at which expression occurs, or in response to external stimuli such as physiological stresses, pathogens, metal ions, or inducing agents. Representative examples of promoters include the bacteriophage T7 promoter, bacteriophage T3 promoter, SP6 promoter, lac operator-promoter, tac promoter, SV40 late promoter, SV40 early promoter, RSV-LTR promoter, CMV IE promoter, EF-1 Alpha promoter, CAG promoter, SV40 early promoter or SV40 late promoter and the CMV IE promoter.

As used throughout the disclosure, the term “substantially complementary” refers to a first sequence that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98% or 99% identical to the complement of a second sequence over a region of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 180, 270, 360, 450, 540, or more nucleotides or amino acids, or that the two sequences hybridize under stringent hybridization conditions.

As used throughout the disclosure, the term “substantially identical” refers to a first and second sequence are at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98% or 99% identical over a region of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 180, 270, 360, 450, 540 or more nucleotides or amino acids, or with respect to nucleic acids, if the first sequence is substantially complementary to the complement of the second sequence.

As used throughout the disclosure, the term “variant” when used to describe a nucleic acid, refers to (i) a portion or fragment of a referenced nucleotide sequence; (ii) the complement of a referenced nucleotide sequence or portion thereof; (iii) a nucleic acid that is substantially identical to a referenced nucleic acid or the complement thereof; or (iv) a nucleic acid that hybridizes under stringent conditions to the referenced nucleic acid, complement thereof, or a sequences substantially identical thereto.

As used throughout the disclosure, the term “vector” refers to a nucleic acid sequence containing an origin of replication. A vector can be a viral vector, bacteriophage, bacterial artificial chromosome or yeast artificial chromosome. A vector can be a DNA or RNA vector. A vector can be a self-replicating extrachromosomal vector, and preferably, is a DNA plasmid. A vector may comprise a combination of an amino acid with a DNA sequence, an RNA sequence, or both a DNA and an RNA sequence.

As used throughout the disclosure, the term “variant” when used to describe a peptide or polypeptide, refers to a peptide or polypeptide that differs in amino acid sequence by the insertion, deletion, or conservative substitution of amino acids, but retain at least one biological activity. Variant can also mean a protein with an amino acid sequence that is substantially identical to a referenced protein with an amino acid sequence that retains at least one biological activity.

A conservative substitution of an amino acid, i.e., replacing an amino acid with a different amino acid of similar properties (e.g., hydrophilicity, degree and distribution of charged regions) is recognized in the art as typically involving a minor change. These minor changes can be identified, in part, by considering the hydropathic index of amino acids, as understood in the art. Kyte et al., J. Mol. Biol. 157: 105-132 (1982). The hydropathic index of an amino acid is based on a consideration of its hydrophobicity and charge. Amino acids of similar hydropathic indexes can be substituted and still retain protein function. In one aspect, amino acids having hydropathic indexes of ±2 are substituted. The hydrophilicity of amino acids can also be used to reveal substitutions that would result in proteins retaining biological function. A consideration of the hydrophilicity of amino acids in the context of a peptide permits calculation of the greatest local average hydrophilicity of that peptide, a useful measure that has been reported to correlate well with antigenicity and immunogenicity. U.S. Pat. No. 4,554,101, incorporated fully herein by reference.

Substitution of amino acids having similar hydrophilicity values can result in peptides retaining biological activity, for example immunogenicity. Substitutions can be performed with amino acids having hydrophilicity values within ±2 of each other. Both the hyrophobicity index and the hydrophilicity value of amino acids are influenced by the particular side chain of that amino acid. Consistent with that observation, amino acid substitutions that are compatible with biological function are understood to depend on the relative similarity of the amino acids, and particularly the side chains of those amino acids, as revealed by the hydrophobicity, hydrophilicity, charge, size, and other properties.

As used herein, “conservative” amino acid substitutions may be defined as set out in Tables A, B, or C below. In some embodiments, fusion polypeptides and/or nucleic acids encoding such fusion polypeptides include conservative substitutions have been introduced by modification of polynucleotides encoding polypeptides of the invention. Amino acids can be classified according to physical properties and contribution to secondary and tertiary protein structure. A conservative substitution is a substitution of one amino acid for another amino acid that has similar properties. Exemplary conservative substitutions are set out in Table A.

TABLE A

Conservative Substitutions I

Side chain characteristics
Amino Acid

Aliphatic
Non-polar
G A P I L V F

Polar-uncharged
C S T M N Q

Polar-charged
D E K R

Aromatic

H F W Y

Other

N Q D E

Alternately, conservative amino acids can be grouped as described in Lehninger, (Biochemistry, Second Edition; Worth Publishers, Inc. NY, N.Y. (1975), pp. 71-77) as set forth in Table B.

TABLE B

Conservative Substitutions II

Side Chain Characteristic
Amino Acid

Non-polar
Aliphatic:
A L I V P

(hydrophobic)
Aromatic:
F W Y

Sulfur-containing:
M

Borderline:
G Y

Uncharged-polar
Hydroxyl:
S T Y

Amides:
N Q

Sulfhydryl:
C

Borderline:
G Y

Positively Charged (Basic):
K R H

Negatively Charged (Acidic):
D E

Alternately, exemplary conservative substitutions are set out in Table C.

TABLE C

Conservative Substitutions III

Original Residue
Exemplary Substitution

Ala (A)
Val Leu Ile Met

Arg (R)
Lys His

Asn (N)
Gln

Asp (D)
Glu

Cys (C)
Ser Thr

Gln (Q)
Asn

Glu (E)
Asp

Gly (G)
Ala Val Leu Pro

His (H)
Lys Arg

Ile (I)
Leu Val Met Ala Phe

Leu (L)
Ile Val Met Ala Phe

Lys (K)
Arg His

Met (M)
Leu Ile Val Ala

Phe (F)
Trp Tyr Ile

Pro (P)
Gly Ala Val Leu Ile

Ser (S)
Thr

Thr (T)
Ser

Trp (W)
Tyr Phe Ile

Tyr (Y)
Trp Phe Thr Ser

Val (V)
Ile Leu Met Ala

It should be understood that the polypeptides of the disclosure are intended to include polypeptides bearing one or more insertions, deletions, or substitutions, or any combination thereof, of amino acid residues as well as modifications other than insertions, deletions, or substitutions of amino acid residues. Polypeptides or nucleic acids of the disclosure may contain one or more conservative substitution.

As used throughout the disclosure, the term “more than one” of the aforementioned amino acid substitutions refers to 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more of the recited amino acid substitutions. The term “more than one” may refer to 2, 3, 4, or 5 of the recited amino acid substitutions.

Polypeptides and proteins of the disclosure, either their entire sequence, or any portion thereof, may be non-naturally occurring. Polypeptides and proteins of the disclosure may contain one or more mutations, substitutions, deletions, or insertions that do not naturally-occur, rendering the entire amino acid sequence non-naturally occurring. Polypeptides and proteins of the disclosure may contain one or more duplicated, inverted or repeated sequences, the resultant sequence of which does not naturally-occur, rendering the entire amino acid sequence non-naturally occurring. Polypeptides and proteins of the disclosure may contain modified, artificial, or synthetic amino acids that do not naturally-occur, rendering the entire amino acid sequence non-naturally occurring.

As used throughout the disclosure, “sequence identity” may be determined by using the stand-alone executable BLAST engine program for blasting two sequences (bl2seq), which can be retrieved from the National Center for Biotechnology Information (NCBI) ftp site, using the default parameters (Tatusova and Madden, FEMS Microbiol Lett., 1999, 174, 247-250; which is incorporated herein by reference in its entirety). The terms “identical” or “identity” when used in the context of two or more nucleic acids or polypeptide sequences, refer to a specified percentage of residues that are the same over a specified region of each of the sequences. The percentage can be calculated by optimally aligning the two sequences, comparing the two sequences over the specified region, determining the number of positions at which the identical residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the specified region, and multiplying the result by 100 to yield the percentage of sequence identity. In cases where the two sequences are of different lengths or the alignment produces one or more staggered ends and the specified region of comparison includes only a single sequence, the residues of single sequence are included in the denominator but not the numerator of the calculation. When comparing DNA and RNA, thymine (T) and uracil (U) can be considered equivalent. Identity can be performed manually or by using a computer sequence algorithm such as BLAST or BLAST 2.0.

As used throughout the disclosure, the term “endogenous” refers to nucleic acid or protein sequence naturally associated with a target gene or a host cell into which it is introduced.

As used throughout the disclosure, the term “exogenous” refers to nucleic acid or protein sequence not naturally associated with a target gene or a host cell into which it is introduced, including non-naturally occurring multiple copies of a naturally occurring nucleic acid, e.g., DNA sequence, or naturally occurring nucleic acid sequence located in a non-naturally occurring genome location.

The disclosure provides methods of introducing a polynucleotide construct comprising a DNA sequence into a host cell. By “introducing” is intended presenting to the plant the polynucleotide construct in such a manner that the construct gains access to the interior of the host cell. The methods of the invention do not depend on a particular method for introducing a polynucleotide construct into a host cell, only that the polynucleotide construct gains access to the interior of one cell of the host. Methods for introducing polynucleotide constructs into bacteria, plants, fungi and animals are known in the art including, but not limited to, stable transformation methods, transient transformation methods, and virus-mediated methods.

Transposons/Transposases

Exemplary transposon/transposase systems of the disclosure include, but are not limited to, piggyBac transposons and transposases, Sleeping Beauty transposons and transposases, Helraiser transposons and transposases and Tol2 transposons and transposases.

The piggyBac transposase recognizes transposon-specific inverted terminal repeat sequences (ITRs) on the ends of the transposon, and moves the contents between the ITRs into TTAA chromosomal sites. The piggyBac transposon system has no payload limit for the genes of interest that can be included between the ITRs. In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac transposon, the transposase is a piggyBac™ or a Super piggyBac™ (SPB) transposase. In certain embodiments, and, in particular, those embodiments wherein the transposase is a Super piggyBac™ (SPB) transposase, the sequence encoding the transposase is an mRNA sequence.

(SEQ ID NO: 14484)

1
MGSSLDDEHI LSALLQSDDE LVGEDSDSEV SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61
SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121
PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTSATFRD TNEDEIYAFF

181
GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDREDFL IRCLRMDDKS IRPTLRENDV

241
FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RVYIPNKPSK YGIKILMMCD

301
SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ

361
EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC

421
DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481
SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPKEV

541
PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF.

The sleeping beauty transposon is transposed into the target genome by the Sleeping Beauty transposase that recognizes ITRs, and moves the contents between the ITRs into TA chromosomal sites. In various embodiments, SB transposon-mediated gene transfer, or gene transfer using any of a number of similar transposons, may be used in the compositions and methods of the disclosure.

In certain embodiments, and, in particular, those embodiments wherein the transposon is a Sleeping Beauty transposon, the transposase is a Sleeping Beauty transposase or a hyperactive Sleeping Beauty transposase (SB100X).

In certain embodiments of the methods of the disclosure, the Sleeping Beauty transposase enzyme comprises an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14485)

1
MGKSKEISQD LRKKIVDLHK
SGSSLGAISK RLKVPRSSVQ TIVRKYKHHG TTQPSYRSGR

61
RRVLSPRDER TLVRKVQINP
RTTAKDLVKM LEETGTKVSI STVKRVLYRH NLKGRSARKK

121
PLLQNRHKKA RLRFATAHGD
KDRTFWRNVL WSDETKIELF GHNDHRYVWR KKGEACKPKN

181
TIPTVKHGGG SIMLWGCFAA
GGTGALHKID GIMRKENYVD ILKQHLKTSV RKLKLGRKWV

241
FQMDNDPKHT SKVVAKWLKD
NKVKVLEWPS QSPDLNPIEN LWAELKKRVR ARRPTNLTQL

301
HQLCQEEWAK IHPTYCGKLV
EGYPKRLTQV KQFKGNATKY.

In certain embodiments of the methods of the disclosure, the hyperactive Sleeping Beauty (SB100X) transposase enzyme comprises an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

The Helraiser transposon is transposed by the Helitron transposase. Helitron transposases mobilize the Helraiser transposon, an ancient element from the bat genome that was active about 30 to 36 million years ago. An exemplary Helraiser transposon of the disclosure includes Helibat1, which comprises a nucleic acid sequence comprising:

(SEQ ID NO: 14652)

1
TCCTATATAA TAAAAGAGAA ACATGCAAAT TGACCATCCC TCCGCTACGC TCAAGCCACG

61
CCCACCAGCC AATCAGAAGT GACTATGCAA ATTAACCCAA CAAAGATGGC AGTTAAATTT

121
GCATACGCAG GTGTCAAGCG CCCCAGGAGG CAACGGCGGC CGCGGGCTCC CAGGACCTTC

181
GCTGGCCCCG GGAGGCGAGG CCGGCCGCGC CTAGCCACAC CCGCGGGCTC CCGGGACCTT

241
CGCCAGCAGA GAGCAGAGCG GGAGAGCGGG CGGAGAGCGG GAGGTTTGGA GGACTTGGCA

301
GAGCAGGAGG CCGCTGGACA TAGAGCAGAG CGAGAGAGAG GGTGGCTTGG AGGGCGTGGC

361
TCCCTCTGTC ACCCCAGCTT CCTCATCACA GCTGTGGAAA CTGACAGCAG GGAGGAGGAA

421
GTCCCACCCC CACAGAATCA GCCAGAATCA GCCGTTGGTC AGACAGCTCT CAGCGGCCTG

481
ACAGCCAGGA CTCTCATTCA CCTGCATCTC AGACCGTGAC AGTAGAGAGG TGGGACTATG

541
TCTAAAGAAC AACTGTTGAT ACAACGTAGC TCTGCAGCCG AAAGATGCCG GCGTTATCGA

601
CAGAAAATGT CTGCAGAGCA ACGTGCGTCT GATCTTGAAA GAAGGCGGCG CCTGCAACAG

661
AATGTATCTG AAGAGCAGCT ACTGGAAAAA CGTCGCTCTG AAGCCGAAAA ACAGCGGCGT

721
CATCGACAGA AAATGTCTAA AGACCAACGT GCCTTTGAAG TTGAAAGAAG GCGGTGGCGA

781
CGACAGAATA TGTCTAGAGA ACAGTCATCA ACAAGTACTA CCAATACCGG TAGGAACTGC

841
CTTCTCAGCA AAAATGGAGT ACATGAGGAT GCAATTCTCG AACATAGTTG TGGTGGAATG

901
ACTGTTCGAT GTGAATTTTG CCTATCACTA AATTTCTCTG ATGAAAAACC ATCCGATGGG

961
AAATTTACTC GATGTTGTAG CAAAGGGAAA GTCTGTCCAA ATGATATACA TTTTCCAGAT

1021
TACCCGGCAT ATTTAAAAAG ATTAATGACA AACGAAGATT CTGACAGTAA AAATTTCATG

1081
GAAAATATTC GTTCCATAAA TAGTTCTTTT GCTTTTGCTT CCATGGGTGC AAATATTGCA

1141
TCGCCATCAG GATATGGGCC ATACTGTTTT AGAATACACG GACAAGTTTA TCACCGTACT

1201
GGAACTTTAC ATCCTTCGGA TGGTGTTTCT CGGAAGTTTG CTCAACTCTA TATTTTGGAT

1261
ACAGCCGAAG CTACAAGTAA AAGATTAGCA ATGCCAGAAA ACCAGGGCTG CTCAGAAAGA

1321
CTCATGATCA ACATCAACAA CCTCATGCAT GAAATAAATG AATTAACAAA ATCGTACAAG

1381
ATGCTACATG AGGTAGAAAA GGAAGCCCAA TCTGAAGCAG CAGCAAAAGG TATTGCTCCC

1441
ACAGAAGTAA CAATGGCGAT TAAATACGAT CGTAACAGTG ACCCAGGTAG ATATAATTCT

1501
CCCCGTGTAA CCGAGGTTGC TGTCATATTC AGAAACGAAG ATGGAGAACC TCCTTTTGAA

1561
AGGGACTTGC TCATTCATTG TAAACCAGAT CCCAATAATC CAAATGCCAC TAAAATGAAA

1621
CAAATCAGTA TCCTGTTTCC TACATTAGAT GCAATGACAT ATCCTATTCT TTTTCCACAT

1681
GGTGAAAAAG GCTGGGGAAC AGATATTGCA TTAAGACTCA GAGACAACAG TGTAATCGAC

1741
AATAATACTA GACAAAATGT AAGGACACGA GTCACACAAA TGCAGTATTA TGGATTTCAT

1801
CTCTCTGTGC GGGACACGTT CAATCCTATT TTAAATGCAG GAAAATTAAC TCAACAGTTT

1861
ATTGTGGATT CATATTCAAA AATGGAGGCC AATCGGATAA ATTTCATCAA AGCAAACCAA

1921
TCTAAGTTGA GAGTTGAAAA ATATAGTGGT TTGATGGATT ATCTCAAATC TAGATCTGAA

1981
AATGACAATG TGCCGATTGG TAAAATGATA ATACTTCCAT CATCTTTTGA GGGTAGTCCC

2041
AGAAATATGC AGCAGCGATA TCAGGATGCT ATGGCAATTG TAACGAAGTA TGGCAAGCCC

2101
GATTTATTCA TAACCATGAC ATGCAACCCC AAATGGGCAG ATATTACAAA CAATTTACAA

2161
CGCTGGCAAA AAGTTGAAAA CAGACCTGAC TTGGTAGCCA GAGTTTTTAA TATTAAGCTG

2221
AATGCTCTTT TAAATGATAT ATGTAAATTC CATTTATTTG GCAAAGTAAT AGCTAAAATT

2281
CATGTCATTG AATTTCAGAA ACGCGGACTG CCTCACGCTC ACATATTATT GATATTAGAT

2341
AGTGAGTCCA AATTACGTTC AGAAGATGAC ATTGACCGTA TAGTTAAGGC AGAAATTCCA

2401
GATGAAGACC AGTGTCCTCG ACTTTTTCAA ATTGTAAAAT CAAATATGGT ACATGGACCA

2461
TGTGGAATAC AAAATCCAAA TAGTCCATGT ATGGAAAATG GAAAATGTTC AAAGGGATAT

2521
CCAAAAGAAT TTCAAAATGC GACCATTGGA AATATTGATG GATATCCCAA ATACAAACGA

2581
AGATCTGGTA GCACCATGTC TATTGGAAAT AAAGTTGTCG ATAACACTTG GATTGTCCCT

2641
TATAACCCGT ATTTGTGCCT TAAATATAAC TGTCATATAA ATGTTGAAGT CTGTGCATCA

2701
ATTAAAAGTG TCAAATATTT ATTTAAATAC ATCTATAAAG GGCACGATTG TGCAAATATT

2761
CAAATTTCTG AAAAAAATAT TATCAATCAT GACGAAGTAC AGGACTTCAT TGACTCCAGG

2821
TATGTGAGCG CTCCTGAGGC TGTTTGGAGA CTTTTTGCAA TGCGAATGCA TGACCAATCT

2881
CATGCAATCA CAAGATTAGC TATTCATTTG CCAAATGATC AGAATTTGTA TTTTCATACC

2941
GATGATTTTG CTGAAGTTTT AGATAGGGCT AAAAGGCATA ACTCGACTTT GATGGCTTGG

3001
TTCTTATTGA ATAGAGAAGA TTCTGATGCA CGTAATTATT ATTATTGGGA GATTCCACAG

3061
CATTATGTGT TTAATAATTC TTTGTGGACA AAACGCCGAA AGGGTGGGAA TAAAGTATTA

3121
GGTAGACTGT TCACTGTGAG CTTTAGAGAA CCAGAACGAT ATTACCTTAG ACTTTTGCTT

3181
CTGCATGTAA AAGGTGCGAT AAGTTTTGAG GATCTGCGAA CTGTAGGAGG TGTAACTTAT

3241
GATACATTTC ATGAAGCTGC TAAACACCGA GGATTATTAC TTGATGACAC TATCTGGAAA

3301
GATACGATTG ACGATGCAAT CATCCTTAAT ATGCCCAAAC AACTACGGCA ACTTTTTGCA

3361
TATATATGTG TGTTTGGATG TCCTTCTGCT GCAGACAAAT TATGGGATGA GAATAAATCT

3421
CATTTTATTG AAGATTTCTG TTGGAAATTA CACCGAAGAG AAGGTGCCTG TGTGAACTGT

3481
GAAATGCATG CCCTTAACGA AATTCAGGAG GTATTCACAT TGCATGGAAT GAAATGTTCA

3541
CATTTCAAAC TTCCGGACTA TCCTTTATTA ATGAATGCAA ATACATGTGA TCAATTGTAC

3601
GAGCAACAAC AGGCAGAGGT TTTGATAAAT TCTCTGAATG ATGAACAGTT GGCAGCCTTT

3661
CAGACTATAA CTTCAGCCAT CGAAGATCAA ACTGTACACC CCAAATGCTT TTTCTTGGAT

3721
GGTCCAGGTG GTAGTGGAAA AACATATCTG TATAAAGTTT TAACACATTA TATTAGAGGT

3781
CGTGGTGGTA CTGTTTTACC CACAGCATCT ACAGGAATTG CTGCAAATTT ACTTCTTGGT

3841
GGAAGAACCT TTCATTCCCA ATATAAATTA CCAATTCCAT TAAATGAAAC TTCAATTTCT

3901
AGACTCGATA TAAAGAGTGA AGTTGCTAAA ACCATTAAAA AGGCCCAACT TCTCATTATT

3961
GATGAATGCA CCATGGCATC CAGTCATGCT ATAAACGCCA TAGATAGATT ACTAAGAGAA

4021
ATTATGAATT TGAATGTTGC ATTTGGTGGG AAAGTTCTCC TTCTCGGAGG GGATTTTCGA

4081
CAATGTCTCA GTATTGTACC ACATGCTATG CGATCGGCCA TAGTACAAAC GAGTTTAAAG

4141
TACTGTAATG TTTGGGGATG TTTCAGAAAG TTGTCTCTTA AAACAAATAT GAGATCAGAG

4201
GATTCTGCTT ATAGTGAATG GTTAGTAAAA CTTGGAGATG GCAAACTTGA TAGCAGTTTT

4261
CATTTAGGAA TGGATATTAT TGAAATCCCC CATGAAATGA TTTGTAACGG ATCTATTATT

4321
GAAGCTACCT TTGGAAATAG TATATCTATA GATAATATTA AAAATATATC TAAACGTGCA

4381
ATTCTTTGTC CAAAAAATGA GCATGTTCAA AAATTAAATG AAGAAATTTT GGATATACTT

4441
GATGGAGATT TTCACACATA TTTGAGTGAT GATTCCATTG ATTCAACAGA TGATGCTGAA

4501
AAGGAAAATT TTCCCATCGA ATTTCTTAAT AGTATTACTC CTTCGGGAAT GCCGTGTCAT

4561
AAATTAAAAT TGAAAGTGGG TGCAATCATC ATGCTATTGA GAAATCTTAA TAGTAAATGG

4621
GGTCTTTGTA ATGGTACTAG ATTTATTATC AAAAGATTAC GACCTAACAT TATCGAAGCT

4681
GAAGTATTAA CAGGATCTGC AGAGGGAGAG GTTGTTCTGA TTCCAAGAAT TGATTTGTCC

4741
CCATCTGACA CTGGCCTCCC ATTTAAATTA ATTCGAAGAC AGTTTCCCGT GATGCCAGCA

4801
TTTGCGATGA CTATTAATAA ATCACAAGGA CAAACTCTAG ACAGAGTAGG AATATTCCTA

4861
CCTGAACCCG TTTTCGCACA TGGTCAGTTA TATGTTGCTT TCTCTCGAGT TCGAAGAGCA

4921
TGTGACGTTA AAGTTAAAGT TGTAAATACT TCATCACAAG GGAAATTAGT CAAGCACTCT

4981
GAAAGTGTTT TTACTCTTAA TGTGGTATAC AGGGAGATAT TAGAATAAGT TTAATCACTT

5041
TATCAGTCAT TGTTTGCATC AATGTTGTTT TTATATCATG TTTTTGTTGT TTTTATATCA

5101
TGTCTTTGTT GTTGTTATAT CATGTTGTTA TTGTTTATTT ATTAATAAAT TTATGTATTA

5161
TTTTCATATA CATTTTACTC ATTTCCTTTC ATCTCTCACA CTTCTATTAT AGAGAAAGGG

5221
CAAATAGCAA TATTAAAATA TTTCCTCTAA TTAATTCCCT TTCAATGTGC ACGAATTTCG

5281
TGCACCGGGC CACTAG.

Unlike other transposases, the Helitron transposase does not contain an RNase-H like catalytic domain, but instead comprises a RepHel motif made up of a replication initiator domain (Rep) and a DNA helicase domain. The Rep domain is a nuclease domain of the HUH superfamily of nucleases.

An exemplary Helitron transposase of the disclosure comprises an amino acid sequence comprising:

In Helitron transpositions, a hairpin close to the 3′ end of the transposon functions as a terminator. However, this hairpin can be bypassed by the transposase, resulting in the transduction of flanking sequences. In addition, Helraiser transposition generates covalently closed circular intermediates. Furthermore, Helitron transpositions can lack target site duplications. In the Helraiser sequence, the transposase is flanked by left and right terminal sequences termed LTS and RTS. These sequences terminate with a conserved 5′-TC/CTAG-3′ motif. A 19 bp palindromic sequence with the potential to form the hairpin termination structure is located 11 nucleotides upstream of the RTS and consists of the sequence

(SEQ ID NO: 14500)

GTGCACGAATTTCGTGCACCGGGCCACTAG.

Tol2 transposons may be isolated or derived from the genome of the medaka fish, and may be similar to transposons of the hAT family. Exemplary Tol2 transposons of the disclosure are encoded by a sequence comprising about 4.7 kilobases and contain a gene encoding the Tol2 transposase, which contains four exons. An exemplary Tol2 transposase of the disclosure comprises an amino acid sequence comprising the following:

(SEQ ID NO: 14502)

1
MEEVCDSSAA ASSTVQNQPQ DQEHPWPYLR EFFSLSGVNK DSFKMKCVLC LPLNKEISAF

61
KSSPSNLRKH IERMHPNYLK NYSKLTAQKR KIGTSTHASS SKQLKVDSVF PVKHVSPVTV

121
NKAILRYIIQ GLHPFSTVDL PSFKELISTL QPGISVITRP TLRSKIAEAA LIMKQKVTAA

181
MSEVEWIATT TDCWTARRKS FIGVTAHWIN PGSLERHSAA LACKRLMGSH TFEVLASAMN

241
DIHSEYEIRD KVVCTTTDSG SNFMKAFRVF GVENNDIETE ARRCESDDTD SEGCGEGSDG

301
VEFQDASRVL DQDDGFEFQL PKHQKCACHL LNLVSSVDAQ KALSNEHYKK LYRSVFGKCQ

361
ALWNKSSRSA LAAEAVESES RLQLLRPNQT RWNSTFMAVD RILQICKEAG EGALRNICTS

421
LEVPMFNPAE MLFLTEWANT MRPVAKVLDI LQAETNTQLG WLLPSVHQLS LKLQRLHHSL

481
RYCDPLVDAL QQGIQTRFKH MFEDPEITAA AILLPKFRTS WTNDETIIKR GMDYIRVHLE

541
PLDHKKELAN SSSDDEDFFA SLKPTTHEAS KELDGYLACV SDTRESLLTF PAICSLSIKT

601
NTPLPASAAC ERLFSTAGLL FSPKRARLDT NNFENQLLLK LNLRFYNFE.

An exemplary Tol2 transposon of the disclosure, including inverted repeats, subterminal sequences and the Tol2 transposase, is encoded by a nucleic acid sequence comprising the following:

(SEQ ID NO: 14653)

1
CAGAGGTGTA AAGTACTTGA GTAATTTTAC TTGATTACTG TACTTAAGTA TTATTTTTGG

61
GGATTTTTAC TTTACTTGAG TACAATTAAA AATCAATACT TTTACTTTTA CTTAATTACA

121
TTTTTTTAGA AAAAAAAGTA CTTTTTACTC CTTACAATTT TATTTACAGT CAAAAAGTAC

181
TTATTTTTTG GAGATCACTT CATTCTATTT TCCCTTGCTA TTACCAAACC AATTGAATTG

241
CGCTGATGCC CAGTTTAATT TAAATGTTAT TTATTCTGCC TATGAAAATC GTTTTCACAT

301
TATATGAAAT TGGTCAGACA TGTTCATTGG TCCTTTGGAA GTGACGTCAT GTCACATCTA

361
TTACCACAAT GCACAGCACC TTGACCTGGA AATTAGGGAA ATTATAACAG TCAATCAGTG

421
GAAGAAAATG GAGGAAGTAT GTGATTCATC AGCAGCTGCG AGCAGCACAG TCCAAAATCA

481
GCCACAGGAT CAAGAGCACC CGTGGCCGTA TCTTCGCGAA TTCTTTTCTT TAAGTGGTGT

541
AAATAAAGAT TCATTCAAGA TGAAATGTGT CCTCTGTCTC CCGCTTAATA AAGAAATATC

601
GGCCTTCAAA AGTTCGCCAT CAAACCTAAG GAAGCATATT GAGGTAAGTA CATTAAGTAT

661
TTTGTTTTAC TGATAGTTTT TTTTTTTTTT TTTTTTTTTT TTTTTGGGTG TGCATGTTTT

721
GACGTTGATG GCGCGCCTTT TATATGTGTA GTAGGCCTAT TTTCACTAAT GCATGCGATT

781
GACAATATAA GGCTCACGTA ATAAAATGCT AAAATGCATT TGTAATTGGT AACGTTAGGT

841
CCACGGGAAA TTTGGCGCCT ATTGCAGCTT TGAATAATCA TTATCATTCC GTGCTCTCAT

901
TGTGTTTGAA TTCATGCAAA ACACAAGAAA ACCAAGCGAG AAATTTTTTT CCAAACATGT

961
TGTATTGTCA AAACGGTAAC ACTTTACAAT GAGGTTGATT AGTTCATGTA TTAACTAACA

1021
TTAAATAACC ATGAGCAATA CATTTGTTAC TGTATCTGTT AATCTTTGTT AACGTTAGTT

1081
AATAGAAATA CAGATGTTCA TTGTTTGTTC ATGTTAGTTC ACAGTGCATT AACTAATGTT

1141
AACAAGATAT AAAGTATTAG TAAATGTTGA AATTAACATG TATACGTGCA GTTCATTATT

1201
AGTTCATGTT AACTAATGTA GTTAACTAAC GAACCTTATT GTAAAAGTGT TACCATCAAA

1261
ACTAATGTAA TGAAATCAAT TCACCCTGTC ATGTCAGCCT TACAGTCCTG TGTTTTTGTC

1321
AATATAATCA GAAATAAAAT TAATGTTTGA TTGTCACTAA ATGCTACTGT ATTTCTAAAA

1381
TCAACAAGTA TTTAACATTA TAAAGTGTGC AATTGGCTGC AAATGTCAGT TTTATTAAAG

1441
GGTTAGTTCA CCCAAAAATG AAAATAATGT CATTAATGAC TCGCCCTCAT GTCGTTCCAA

1501
GCCCGTAAGA CCTCCGTTCA TCTTCAGAAC ACAGTTTAAG ATATTTTAGA TTTAGTCCGA

1561
GAGCTTTCTG TGCCTCCATT GAGAATGTAT GTACGGTATA CTGTCCATGT CCAGAAAGGT

1621
AATAAAAACA TCAAAGTAGT CCATGTGACA TCAGTGGGTT AGTTAGAATT TTTTGAAGCA

1681
TCGAATACAT TTTGGTCCAA AAATAACAAA ACCTACGACT TTATTCGGCA TTGTATTCTC

1741
TTCCGGGTCT GTTGTCAATC CGCGTTCACG ACTTCGCAGT GACGCTACAA TGCTGAATAA

1801
AGTCGTAGGT TTTGTTATTT TTGGACCAAA ATGTATTTTC GATGCTTCAA ATAATTCTAC

1861
CTAACCCACT GATGTCACAT GGACTACTTT GATGTTTTTA TTACCTTTCT GGACATGGAC

1921
AGTATACCGT ACATACATTT TCAGTGGAGG GACAGAAAGC TCTCGGACTA AATCTAAAAT

1981
ATCTTAAACT GTGTTCCGAA GATGAACGGA GGTGTTACGG GCTTGGAACG ACATGAGGGT

2041
GAGTCATTAA TGACATCTTT TCATTTTTGG GTGAACTAAC CCTTTAATGC TGTAATCAGA

2101
GAGTGTATGT GTAATTGTTA CATTTATTGC ATACAATATA AATATTTATT TGTTGTTTTT

2161
ACAGAGAATG CACCCAAATT ACCTCAAAAA CTACTCTAAA TTGACAGCAC AGAAGAGAAA

2221
GATCGGGACC TCCACCCATG CTTCCAGCAG TAAGCAACTG AAAGTTGACT CAGTTTTCCC

2281
AGTCAAACAT GTGTCTCCAG TCACTGTGAA CAAAGCTATA TTAAGGTACA TCATTCAAGG

2341
ACTTCATCCT TTCAGCACTG TTGATCTGCC ATCATTTAAA GAGCTGATTA GTACACTGCA

2401
GCCTGGCATT TCTGTCATTA CAAGGCCTAC TTTACGCTCC AAGATAGCTG AAGCTGCTCT

2461
GATCATGAAA CAGAAAGTGA CTGCTGCCAT GAGTGAAGTT GAATGGATTG CAACCACAAC

2521
GGATTGTTGG ACTGCACGTA GAAAGTCATT CATTGGTGTA ACTGCTCACT GGATCAACCC

2581
TGGAAGTCTT GAAAGACATT CCGCTGCACT TGCCTGCAAA AGATTAATGG GCTCTCATAC

2641
TTTTGAGGTA CTGGCCAGTG CCATGAATGA TATCCACTCA GAGTATGAAA TACGTGACAA

2701
GGTTGTTTGC ACAACCACAG ACAGTGGTTC CAACTTTATG AAGGCTTTCA GAGTTTTTGG

2761
TGTGGAAAAC AATGATATCG AGACTGAGGC AAGAAGGTGT GAAAGTGATG ACACTGATTC

2821
TGAAGGCTGT GGTGAGGGAA GTGATGGTGT GGAATTCCAA GATGCCTCAC GAGTCCTGGA

2881
CCAAGACGAT GGCTTCGAAT TCCAGCTACC AAAACATCAA AAGTGTGCCT GTCACTTACT

2941
TAACCTAGTC TCAAGCGTTG ATGCCCAAAA AGCTCTCTCA AATGAACACT ACAAGAAACT

3001
CTACAGATCT GTCTTTGGCA AATGCCAAGC TTTATGGAAT AAAAGCAGCC GATCGGCTCT

3061
AGCAGCTGAA GCTGTTGAAT CAGAAAGCCG GCTTCAGCTT TTAAGGCCAA ACCAAACGCG

3121
GTGGAATTCA ACTTTTATGG CTGTTGACAG AATTCTTCAA ATTTGCAAAG AAGCAGGAGA

3181
AGGCGCACTT CGGAATATAT GCACCTCTCT TGAGGTTCCA ATGTAAGTGT TTTTCCCCTC

3241
TATCGATGTA AACAAATGTG GGTTGTTTTT GTTTAATACT CTTTGATTAT GCTGATTTCT

3301
CCTGTAGGTT TAATCCAGCA GAAATGCTGT TCTTGACAGA GTGGGCCAAC ACAATGCGTC

3361
CAGTTGCAAA AGTACTCGAC ATCTTGCAAG CGGAAACGAA TACACAGCTG GGGTGGCTGC

3421
TGCCTAGTGT CCATCAGTTA AGCTTGAAAC TTCAGCGACT CCACCATTCT CTCAGGTACT

3481
GTGACCCACT TGTGGATGCC CTACAACAAG GAATCCAAAC ACGATTCAAG CATATGTTTG

3541
AAGATCCTGA GATCATAGCA GCTGCCATCC TTCTCCCTAA ATTTCGGACC TCTTGGACAA

3601
ATGATGAAAC CATCATAAAA CGAGGTAAAT GAATGCAAGC AACATACACT TGACGAATTC

3661
TAATCTGGGC AACCTTTGAG CCATACCAAA ATTATTCTTT TATTTATTTA TTTTTGCACT

3721
TTTTAGGAAT GTTATATCCC ATCTTTGGCT GTGATCTCAA TATGAATATT GATGTAAAGT

3781
ATTCTTGCAG CAGGTTGTAG TTATCCCTCA GTGTTTCTTG AAACCAAACT CATATGTATC

3841
ATATGTGGTT TGGAAATGCA GTTAGATTTT ATGCTAAAAT AAGGGATTTG CATGATTTTA

3901
GATGTAGATG ACTGCACGTA AATGTAGTTA ATGACAAAAT CCATAAAATT TGTTCCCAGT

3961
CAGAAGCCCC TCAACCAAAC TTTTCTTTGT GTCTGCTCAC TGTGCTTGTA GGCATGGACT

4021
ACATCAGAGT GCATCTGGAG CCTTTGGACC ACAAGAAGGA ATTGGCCAAC AGTTCATCTG

4081
ATGATGAAGA TTTTTTCGCT TCTTTGAAAC CGACAACACA TGAAGCCAGC AAAGAGTTGG

4141
ATGGATATCT GGCCTGTGTT TCAGACACCA GGGAGTCTCT GCTCACGTTT CCTGCTATTT

4201
GCAGCCTCTC TATCAAGACT AATACACCTC TTCCCGCATC GGCTGCCTGT GAGAGGCTTT

4261
TCAGCACTGC AGGATTGCTT TTCAGCCCCA AAAGAGCTAG GCTTGACACT AACAATTTTG

4321
AGAATCAGCT TCTACTGAAG TTAAATCTGA GGTTTTACAA CTTTGAGTAG CGTGTACTGG

4381
CATTAGATTG TCTGTCTTAT AGTTTGATAA TTAAATACAA ACAGTTCTAA AGCAGGATAA

4441
AACCTTGTAT GCATTTCATT TAATGTTTTT TGAGATTAAA AGCTTAAACA AGAATCTCTA

4501
GTTTTCTTTC TTGCTTTTAC TTTTACTTCC TTAATACTCA AGTACAATTT TAATGGAGTA

4561
CTTTTTTACT TTTACTCAAG TAAGATTCTA GCCAGATACT TTTACTTTTA ATTGAGTAAA

4621
ATTTTCCCTA AGTACTTGTA CTTTCACTTG AGTAAAATTT TTGAGTACTT TTTACACCTC

4681
TG.

Exemplary transposon/transposase systems of the disclosure include, but are not limited to, piggyBac and piggyBac-like transposons and transposases.

PiggyBac and piggyBac-like transposases recognizes transposon-specific inverted terminal repeat sequences (ITRs) on the ends of the transposon, and moves the contents between the ITRs into TTAA or TTAT chromosomal sites. The piggyBac or piggyBac-like transposon system has no payload limit for the genes of interest that can be included between the ITRs.

In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac transposon, the transposase is a piggyBac™, Super piggyBac™ (SPB) transposase. In certain embodiments, and, in particular, those embodiments wherein the transposase is a piggyBac™, Super piggyBac™ (SPB), the sequence encoding the transposase is an mRNA sequence.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or a piggyBac-like transposase enzyme. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at one or more of positions 30, 165, 282, or 538 of the sequence:

In certain embodiments, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at two or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at three or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at each of the following positions 30, 165, 282, and 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the amino acid substitution at position 30 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 165 of the sequence of SEQ ID NO: 14487 is a substitution of a serine (S) for a glycine (G). In certain embodiments, the amino acid substitution at position 282 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 538 of the sequence of SEQ ID NO: 14487 is a substitution of a lysine (K) for an asparagine (N).

In certain embodiments of the methods of the disclosure, the transposase enzyme is a Super piggyBac™ (SPB) or piggyBac-like transposase enzyme. In certain embodiments, the Super piggyBac™ (SPB) or piggyBac-like transposase enzyme of the disclosure may comprise or consist of the amino acid sequence of the sequence of SEQ ID NO: 14487 wherein the amino acid substitution at position 30 is a substitution of a valine (V) for an isoleucine (I), the amino acid substitution at position 165 is a substitution of a serine (S) for a glycine (G), the amino acid substitution at position 282 is a substitution of a valine (V) for a methionine (M), and the amino acid substitution at position 538 is a substitution of a lysine (K) for an asparagine (N). In certain embodiments, the Super piggyBac™ (SPB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

In certain embodiments of the methods of the disclosure, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™, Super piggyBac™ or piggyBac-like transposase enzyme may further comprise an amino acid substitution at one or more of positions 3, 46, 82, 103, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 258, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 486, 503, 552, 570 and 591 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™, Super piggyBac™ or piggyBac-like transposase enzyme may further comprise an amino acid substitution at one or more of positions 46, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 485, 503, 552 and 570. In certain embodiments, the amino acid substitution at position 3 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an asparagine (N) for a serine (S). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an alanine (A). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 82 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 103 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a serine (S). In certain embodiments, the amino acid substitution at position 119 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for an arginine (R). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) a cysteine (C). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 185 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 187 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for an alanine (A). In certain embodiments, the amino acid substitution at position 200 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 207 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a valine (V). In certain embodiments, the amino acid substitution at position 209 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a valine (V). In certain embodiments, the amino acid substitution at position 226 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a methionine (M). In certain embodiments, the amino acid substitution at position 235 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a leucine (L). In certain embodiments, the amino acid substitution at position 240 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 241 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 243 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a proline (P). In certain embodiments, the amino acid substitution at position 258 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an asparagine (N). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a proline (P). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine for a proline (P). In certain embodiments, the amino acid substitution at position 315 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for an arginine (R). In certain embodiments, the amino acid substitution at position 319 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a threonine (T). In certain embodiments, the amino acid substitution at position 327 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 328 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a cysteine (C). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 421 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for the aspartic acid (D). In certain embodiments, the amino acid substitution at position 436 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a valine (V). In certain embodiments, the amino acid substitution at position 456 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a methionine (M). In certain embodiments, the amino acid substitution at position 470 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 485 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a serine (S). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a methionine (M). In certain embodiments, the amino acid substitution at position 552 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 570 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a glutamine (Q). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a glutamine (Q).

In certain embodiments of the methods of the disclosure, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or piggyBac-like transposase enzyme or may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments of the methods of the disclosure, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or piggyBac-like transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at two, three, four, five, six or more of positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or piggyBac-like transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, the amino acid substitution at position 103 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a serine (S). In certain embodiments, the amino acid substitution at position 194 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 372 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for an arginine (R). In certain embodiments, the amino acid substitution at position 375 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for a lysine (K). In certain embodiments, the amino acid substitution at position 450 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an asparagine (N) for an aspartic acid (D). In certain embodiments, the amino acid substitution at position 509 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a serine (S). In certain embodiments, the amino acid substitution at position 570 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an asparagine (N). In certain embodiments, the piggyBac™ or piggyBac-like transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487. In certain embodiments, including those embodiments wherein the piggyBac™ or piggyBac-like transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, the piggyBac™ or piggyBac-like transposase enzyme may further comprise an amino acid substitution at positions 372, 375 and 450 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, the piggyBac™ or piggyBac-like transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, a substitution of an alanine (A) for an arginine (R) at position 372 of SEQ ID NO: 14487, and a substitution of an alanine (A) for a lysine (K) at position 375 of SEQ ID NO: 14487. In certain embodiments, the piggyBac™ or piggyBac-like transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, a substitution of an alanine (A) for an arginine (R) at position 372 of SEQ ID NO: 14487, a substitution of an alanine (A) for a lysine (K) at position 375 of SEQ ID NO: 14487 and a substitution of an asparagine (N) for an aspartic acid (D) at position 450 of SEQ ID NO: 14487.

In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from an insect. In certain embodiments, the insect is Trichoplusia ni (GenBank Accession No. AAA87375; SEQ ID NO: 14666), Argyrogramma agnata (GenBank Accession No. GU477713; SEQ ID NO: 14534, SEQ ID NO: 14667), Anopheles gambiae (GenBank Accession No. XP 312615 (SEQ ID NO: 14668); GenBank Accession No. XP 320414 (SEQ ID NO: 14669); GenBank Accession No. XP 310729 (SEQ ID NO: 14670)), Aphis gossypii (GenBank Accession No. GU329918; SEQ ID NO: 14671, SEQ ID NO: 14672), Acyrthosiphon pisum (GenBank Accession No. XP 001948139; SEQ ID NO: 14673), Agrotis ipsilon (GenBank Accession No. GU477714; SEQ ID NO: 14537, SEQ ID NO: 14674), Bombyx mori (GenBank Accession No. BAD11135; SEQ ID NO: 14505), Chilo suppressalis (GenBank Accession No. JX294476; SEQ ID NO: 14675, SEQ ID NO: 14676), Drosophila melanogaster (GenBank Accession No. AAL39784; SEQ ID NO: 14677), Helicoverpa armigera (GenBank Accession No. ABS18391; SEQ ID NO: 14525), Heliothis virescens (GenBank Accession No. ABD76335; SEQ ID NO: 14678), Macdunnoughia crassisigna (GenBank Accession No. EU287451; SEQ ID NO: 14679, SEQ ID NO: 14680), Pectinophora gossypiella (GenBank Accession No. GU270322; SEQ ID NO: 14530, SEQ ID NO: 14681), Tribolium castaneum (GenBank Accession No. XP 001814566; SEQ ID NO: 14682), Ctenoplusia agnata (also called Argyrogramma agnata), Messour bouvieri, Megachile rotundata, Bombus impatiens, Mamestra brassicae, Mayetiola destructor or Apis mellifera.

In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from an insect. In certain embodiments, the insect is Trichoplusia ni (AAA87375).

In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from an insect. In certain embodiments, the insect is Bombyx mori (BAD11135).

In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from a crustacean. In certain embodiments, the crustacean is Daphnia pulicaria (AAM76342, SEQ ID NO: 14683).

In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from a vertebrate. In certain embodiments, the vertebrate is Xenopus tropicalis (GenBank Accession No. BAF82026; SEQ ID NO: 14518), Homo sapiens (GenBank Accession No. NP 689808; SEQ ID NO: 14684), Mus musculus (GenBank Accession No. NP 741958; SEQ ID NO: 14685), Macaca fascicularis (GenBank Accession No. AB179012; SEQ ID NO: 14686, SEQ ID NO: 14687), Rattus norvegicus (GenBank Accession No. XP 220453; SEQ ID NO: 14688) or Myotis lucifugus.

In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from a urochordate. In certain embodiments, the urochordate is Ciona intestinalis (GenBank Accession No. XP 002123602; SEQ ID NO: 14689).

In certain embodiments, the piggyBac or piggyBac-like transposase inserts a transposon at the sequence 5′-TTAT-3′ within a chromosomal site (a TTAT target sequence).

In certain embodiments, the piggyBac or piggyBac-like transposase inserts a transposon at the sequence 5′-TTAA-3′ within a chromosomal site (a TTAA target sequence).

In certain embodiments, the target sequence of the piggyBac or piggyBac-like transposon comprises or consists of 5′-CTAA-3′, 5′-TTAG-3′, 5′-ATAA-3′, 5′-TCAA-3′, 5′AGTT-3′, 5′-ATTA-3′, 5′-GTTA-3′, 5′-TTGA-3′, 5′-TTTA-3′, 5′-TTAC-3′, 5′-ACTA-3′, 5′-AGGG-3′, 5′-CTAG-3′, 5′-TGAA-3′, 5′-AGGT-3′, 5′-ATCA-3′, 5′-CTCC-3′, 5′-TAAA-3′, 5′-TCTC-3′, 5′TGAA-3′, 5′-AAAT-3′, 5′-AATC-3′, 5′-ACAA-3′, 5′-ACAT-3′, 5′-ACTC-3′, 5′-AGTG-3′, 5′-ATAG-3′, 5′-CAAA-3′, 5′-CACA-3′, 5′-CATA-3′, 5′-CCAG-3′, 5′-CCCA-3′, 5′-CGTA-3′, 5′-GTCC-3′, 5′-TAAG-3′, 5′-TCTA-3′, 5′-TGAG-3′, 5′-TGTT-3′, 5′-TTCA-3′5′-TTCT-3′ and 5′-TTTT-3′.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Bombyx mori. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14504)

1
MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61
EANAIIANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121
NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS

181
FYMQETTLCE LKALIALLYL AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLQNN

241
IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ CCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301
IPNKPAKYGI KILALVDAKN FDVVNLEVYA GKQPSGPYAV SNRPFEVVER LIQPVARSHR

361
NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421
VSYAPKKNKV VVVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDVVD ELSANYNVSR

481
NSKRWPMTLF YGVLNMAAIN ACIIYRANKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541
PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KHSCNACAKP ICMEHAKFLC

601
ENCAELDSSL.

The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14505)

1
MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61
EANAIIANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121
NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS

181
FYMQETTLCE LKALIALLYL AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLQNN

241
IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ CCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301
IPNKPAKYGI KILALVDAKN FYVVNLEVYA GKQPSGPYAV SNRPFEVVER LIQPVARSHR

361
NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421
VSYAPKKNKV VVVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDVVD ELCANYNVSR

481
NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541
PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601
ENCAELDSSL.

In certain embodiments, the piggyBac or piggyBac-like transposase is fused to a nuclear localization signal. In certain embodiments, the amino acid sequence of the piggyBac or piggyBac-like transposase fused to a nuclear localization signal is encoded by a polynucleotide sequence comprising:

(SEQ ID NO: 14629)

1
atggcaccca aaaagaaacg taaagtgatg gacattgaaa gacaggaaga aagaatcagg

61
gcgatgctcg aagaagaact gagcgactac tccgacgaat cgtcatcaga ggatgaaacc

121
gaccactgta gcgagcatga ggttaactac gacaccgagg aggagagaat cgactctgtg

181
gatgtgccct ccaactcacg ccaagaagag gccaatgcaa ttatcgcaaa cgaatcggac

241
agcgatccag acgatgatct gccactgtcc ctcgtgcgcc agcgggccag cgcttcgaga

301
caagtgtcag gtccattcta cacttcgaag gacggcacta agtggtacaa gaattgccag

361
cgacctaacg tcagactccg ctccgagaat atcgtgaccg aacaggctca ggtcaagaat

421
atcgcccgcg acgcctcgac tgagtacgag tgttggaata tcttcgtgac ttcggacatg

481
ctgcaagaaa ttctgacgca caccaacagc tcgattaggc atcgccagac caagactgca

541
gcggagaact catcggccga aacctccttc tatatgcaag agactactct gtgcgaactg

601
aaggcgctga ttgcactgct gtacttggcc ggcctcatca aatcaaatag gcagagcctc

661
aaagatctct ggagaacgga tggaactgga gtggatatct ttcggacgac tatgagcttg

721
cagcggttcc agtttctgca aaacaatatc agattcgacg acaagtccac ccgggacgaa

781
aggaaacaga ctgacaacat ggctgcgttc cggtcaatat tcgatcagtt tgtgcagtgc

841
tgccaaaacg cttatagccc atcggaattc ctgaccatcg acgaaatgct tctctccttc

901
cgggggcgct gcctgttccg agtgtacatc ccgaacaagc cggctaaata cggaatcaaa

961
atcctggccc tggtggacgc caagaatttc tacgtcgtga atctcgaagt gtacgcagga

1021
aagcaaccgt cgggaccgta cgctgtttcg aaccgcccgt ttgaagtcgt cgagcggctt

1081
attcagccgg tggccagatc ccaccgcaat gttaccttcg acaattggtt caccggctac

1141
gagctgatgc ttcaccttct gaacgagtac cggctcacta gcgtggggac tgtcaggaag

1201
aacaagcggc agatcccaga atccttcatc cgcaccgacc gccagcctaa ctcgtccgtg

1261
ttcggatttc aaaaggatat cacgcttgtc tcgtacgccc ccaagaaaaa caaggtcgtg

1321
gtcgtgatga gcaccatgca tcacgacaac agcatcgacg agtcaaccgg agaaaagcaa

1381
aagcccgaga tgatcacctt ctacaattca actaaggccg gcgtcgacgt cgtggatgaa

1441
ctgtgcgcga actataacgt gtcccggaac tctaagcggt ggcctatgac tctcttctac

1501
ggagtgctga atatggccgc aatcaacgcg tgcatcatct accgcaccaa caagaacgtg

1561
accatcaagc gcaccgagtt catcagatcg ctgggtttga gcatgatcta cgagcacctc

1621
cattcacgga acaagaagaa gaatatccct acttacctga ggcagcgtat cgagaagcag

1681
ttgggagaac caagcccgcg ccacgtgaac gtgccggggc gctacgtgcg gtgccaagat

1741
tgcccgtaca aaaaggaccg caaaaccaaa agatcgtgta acgcgtgcgc caaacctatc

1801
tgcatggagc atgccaaatt tctgtgtgaa aattgtgctg aactcgattc ctccctg.

In certain embodiments, the piggyBac or piggyBac-like transposase is hyperactive. A hyperactive piggyBac or piggyBac-like transposase is a transposase that is more active than the naturally occurring variant from which it is derived. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase enzyme is isolated or derived from Bombyx mori. In certain embodiments, the piggyBac or piggyBac-like transposase is a hyperactive variant of SEQ ID NO: 14505. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to:

(SEQ ID NO: 14576)

1
MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61
EANAIIANES DSDPDDDLPL SLVRQRASAS RQMSGPHYTS KDGTKWYKNC QRPNVRLRSE

121
NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRWRQTKT AAENSSASTS

181
FYMQETTLCE LKALIGLLYI AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLQNN

241
IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301
IPNKPAKYGI KILALVDAKN FYVKNLEVYA GKQPSGPYAV SNRPFEVVER LIQPVARSHR

361
NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421
VSYAPKKNKV VVVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDVVD ELCANYNVSR

481
NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541
PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601
ENCAELDSHL.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14576. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 14630)

1
MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61
EANAIIANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121
NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRWRQTKT AAENSSAETS

181
FYMQETTLCE LKALIGLLYI AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLLNN

241
IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301
IPNKPAKYGI KILALVDAKN FYVHNLEVYA GKQPSGPYAV SNRPFEVVER LIQPVARSHR

361
NVTFDNWFTG YEVMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421
VSYAPKKNKV VVVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDVVD ELCANYNVSR

481
NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541
PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601
ENCAHLDS.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 14631)

1
MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61
EANAIIANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121
NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRWRQTKT AAENSSASTS

181
FYMQETTLCE LKALIGLLYI AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLLNN

241
IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301
IPNKPAKYGI KILALVDAKN FYVKNLEVYA GKQPSGPYAV SNRPFEVVER LIQPVARSHR

361
NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421
VSYAPKKNKV VVVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDVVD ELCANYNVSR

481
NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541
PTYLRQRIAM QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601
ENCAELDSSL.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 14632)

1
MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61
EANAIIANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121
NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRWRQTKT AAENSSAETS

181
FYMQETTLCE LKALIGLLYI AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLLNN

241
IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301
IPNKPAKYGI KILALVDAKN FYVKNLEVYA GKQPSGPYAV SNRPFEVVER LIQPVARSHR

361
NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKTQIPENF IRTDRQPNSS VFGFQKDITL

421
VSYAPKKNKV VVVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDVVD ELQANYNVSR

481
NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541
PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601
ENCAELDSSL.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 14633)

1
MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61
EANAIIANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121
NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRWRQTKT AAENSSAETS

181
FYMQETTLCE LKALIGLLYI AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLQNN

241
IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301
IPNKPAKYGI KILALVDAKN FYVKNLEVYA GKQPSGPYAV SNRPFEVVER LIQPVARSHR

361
NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421
VSYAPKKNKV VVVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDVVD ELCANYNVSR

481
NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541
PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601
ENCAELDSSL.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 14634)

1
MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61
EANAIIANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121
NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS

181
FYMQETTLCE LKALIALLYL AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLQNN

241
IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ CCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301
IPNKPAKYGI KILALVDAKN DYVVNLEVYA GKQPSGPYAV SNRPFEVVER LIQPVARSHR

361
NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421
VSYAPKKNKV VVVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDVVD ELCANYNVSR

481
NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541
PTYLRQRIEK QLGEPSSRHV NVKGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601
ENCAELDSSL.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase is more active than the transposase of SEQ ID NO: 14505. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% or any percentage in between identical to SEQ ID NO: 14505.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises an amino acid substitution at a position selected from 92, 93, 96, 97, 165, 178, 189, 196, 200, 201, 211, 215, 235, 238, 246, 253, 258, 261, 263, 271, 303, 321, 324, 330, 373, 389, 399, 402, 403, 404, 448, 473, 484, 507, 523, 527, 528, 543, 549, 550, 557, 601, 605, 607, 609, 610 or a combination thereof (relative to SEQ ID NO: 14505). In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises an amino acid substitution of Q92A, V93L, V93M, P96G, F97H, F97C, H165E, H165W, E178S, E178H, C189P, A196G, L200I, A201Q, L211A, W215Y, G2195, Q235Y, Q235G, Q238L, K246I, K253V, M258V, F261L, S263K, C271S, N303R, F321W, F321D, V324K, V324H, A330V, L373C, L373V, V389L, S399N, R402K, T403L, D404Q, D404S, D404M, N441R, G448W, E449A, V469T, C473Q, R484K T507C, G523A, I527M, Y528K Y543I, E549A, K550M, P557S, E601V, E605H, E605W, D607H, 5609H, L610I or any combination thereof. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises an amino acid substitution of Q92A, V93L, V93M, P96G, F97H, F97C, H165E, H165W, E178S, E178H, C189P, A196G, L200I, A201Q, L211A, W215Y, G2195, Q235Y, Q235G, Q238L, K246I, K253V, M258V, F261L, S263K, C271S, N303R, F321W, F321D, V324K, V324H, A330V, L373C, L373V, V389L, S399N, R402K, T403L, D404Q, D404S, D404M, N441R, G448W, E449A, V469T, C473Q, R484K T507C, G523A, I527M, Y528K Y543I, E549A, K550M, P557S, E601V, E605H, E605W, D607H, 5609H and L610I.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises one or more substitutions of an amino acid that is not wild type, wherein the one or more substitutions a for wild type amino acid comprises a substitution of E4X, A12X, M13X, L14X, E15X, D20X, E24X, S25X, S26X, S27X, D32X, H33X, E36X, E44X, E45X, E46X, I48X, D49X, R58X, A62X, N63X, A64X, I65X, I66X, N68X, E69X, D71X, S72X, D76X, P79X, R84X, Q85X, A87X, S88X, Q92X, V93X, S94X, G95X, P96X, F97X, Y98X, T99X, I145X, S149X, D150X, L152X, E154X, T157X, N160X, S161X, S162X, H165X, R166X, T168X, K169X, T170X, A171X, E173X, S175X, S176X, E178X, T179X, M183X, Q184X, T186X, T187X, L188X, C189X, L194X, I195X, A196X, L198X, L200X, A201X, L203X, I204X, K205X, A206X, N207X, Q209X, S210X, L211X, K212X, D213X, L214X, W215X, R216X, T217X, G219X, V222X, D223X, I224X, T227X, M229X, Q235X, L237X, Q238X, N239X, N240X, P302X, N303X, P305X, A306X, K307X, Y308X, I310X, K311X, I312X, L313X, A314X, L315X, V316X, D317X, A318X, K319X, N320X, F321X, Y322X, V323X, V324X, L326X, E327X, V328X, A330X, Q333X, P334X, S335X, G336X, P337X, A339X, V340X, S341X, N342X, R343X, P344X, F345X, E346X, V347X, E349X, I352X, Q353X, V355X, A356X, R357X, N361X, D365X, W367X, T369X, G370X, L373X, M374X, L375X, H376X, N379X, E380X, R382X, V386X, V389X, N392X, R394X, Q395X, S399X, F400X, I401X, R402XT403X, D404X, R405X, Q406X, P407X, N408X, S409X, S410X, V411X, F412X, F414X, Q415X, I418X, T419X, L420X, N428XV432X, M434X, D440X, N441X, S442X, I443X, D444X, E445X, G448X, E449X, Q451X, K452X, M455X, I456X, T457X, F458X, S461X, A464X, V466X, Q468X, V469X, E471X, L472X, C473X, A474X, K483X, W485X, T488X, L489X, Y491X, G492X, V493X, M496X, I499X, C502X, I503X, T507X, K509X, N510X, V511X, T512X, I513X, R515X, E517X, S521X, G523X, L524X, S525X, I527X, Y528X, E529X, H532X, S533X, N535X, K536X, K537X, N539X, I540X, T542X, Y543X, Q546X, E549X, K550X, Q551X, G553X, E554X, P555X, S556X, P557X, R558X, H559X, V560X, N561X, V562X, P563X, G564X, R565X, Y566X, V567X, Q570X, D571X, P573X, Y574X, K576X, K581X, S583X, A586X, A588X, E594X, F598X, L599X, E601X, N602X, C603X, A604X, E605X, L606X, D607X, S608X, S609X or L610X (relative to SEQ ID NO: 14505). A list of hyperactive amino acid substitutions can be found in U.S. Pat. No. 10,041,077, the contents of which are incorporated herein by reference in their entirety.

In certain embodiments, the piggyBac or piggyBac-like transposase is integration deficient. In certain embodiments, an integration deficient piggyBac or piggyBac-like transposase is a transposase that can excise its corresponding transposon, but that integrates the excised transposon at a lower frequency than a corresponding wild type transposase. In certain embodiments, the piggyBac or piggyBac-like transposase is an integration deficient variant of SEQ ID NO: 14505.

In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 14606)

1
MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61
EANAIIANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121
NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS

181
FYMQETTLCE LKALIALLYL AGLIKSNRQS LKDLWRKDGT GVDIFRTTMS LQRFQFLLNN

241
IRFDDISTRD ERKQTDNMAA FRSIFDQFVQ CCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301
IPNKPAKYGI KILALVDAKN FYVVNLEVYA GKQPSGPYAV SNRPFEVVER LIQPVARSHR

361
NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421
VSYAPKKNKV VVVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDVVD ELCANYNVSR

481
NSKKWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMMYEH LHSRNKKKNI

541
PTYLRQRIEK QLGEPVPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601
ENCAELDSSL.

In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 14607)

1
MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61
EANAIIANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121
NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS

181
FYMQETTLCE LKALIGLLYL AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFYFLQNN

241
IRFDDKSTLD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301
IPNKPAKYGI KILALVDAKN FYVVNLEVYA GKQPSGPYAV SNRPFEVVER LIQPVARSHR

361
NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421
VSYAPKKNKV VVVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDVVD ELCANYNVSR

481
NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541
PTYLRQRIEK QLGEPSPRHV NYPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601
VNCAELDSSL.

In certain embodiments, the piggyBac or piggyBac-like transposase that is is integration deficient comprises a sequence of:

(SEQ ID NO: 14608)

1
MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61
EANAIIANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121
NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS

181
FYMQETTLCE LKALIALLYL AGLIKSNRQS LKDLWRKDGT GVDIFRTTMS LQRFQFLLNN

241
IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ CCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301
IPNKPAKYGI KILALVDAKN DYVVNLEVYA GKQPSGPYAV SNRPFEVVER LIQPVARSHR

361
NVTFDNWFTG YECMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421
VSYAPKKNKV VVVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDVVD ELCANYNVSR

481
NSKKWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIKEH LHSRNKKKNI

541
PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601
ENCAELDSSL.

In certain embodiments, the integration deficient transposase comprises a sequence that is at least 90% identical to SEQ ID NO: 14608.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Bombyx mori. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14506)

1
ttatcccggc gagcatgagg cagggtatct cataccctgg taaaatttta aagttgtgta

61
ttttataaaa ttttcgtctg acaacactag cgcgctcagt agctggaggc aggagcgtgc

121
gggaggggat agtggcgtga tcgcagtgtg gcacgggaca ccggcgagat attcgtgtgc

181
aaacctgttt cgggtatgtt ataccctgcc tcattgttga cgtatttttt ttatgtaatt

241
tttccgatta ttaatttcaa ctgttttatt ggtattttta tgttatccat tgttcttttt

301
ttatgattta ctgtatcggt tgtctttcgt tcctttagtt gagttttttt ttattatttt

361
cagtttttga tcaaa.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14507)

1
tcatattttt agtttaaaaa aataattata tgttttataa tgaaaagaat ctcattatct

61
ttcagtatta ggttgattta tattccaaag aataatattt ttgttaaatt gttgattttt

121
gtaaacctct aaatgtttgt tgctaaaatt actgtgttta agaaaaagat taataaataa

181
taataatttc ataattaaaa acttctttca ttgaatgcca ttaaataaac cattatttta

241
caaaataaga tcaacataat tgagtaaata ataataagaa caatattata gtacaacaaa

301
atatgggtat gtcataccct gccacattct tgatgtaact ttttttcacc tcatgctcgc

361
cgggttat.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14508)

1
ttatcccggc gagcatgagg cagggtatct cataccctgg taaaatttta aagttgtgta

61
ttttataaaa ttttcgtctg acaacactag cgcgctcagt agctggaggc aggagcgtgc

121
gggaggggat agtggcgtga tcgcagtgtg gcacgggaca ccggcgagat attcgtgtgc

181
aaacctgttt cgggtatgtt ataccctgcc tcat.

In certain embodiments, the piggyBac™ (PB) or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14509)

1
taaataataa taatttcata attaaaaact tctttcattg aatgccatta aataaaccat

61
tattttacaa aataagatca acataattga gtaaataata ataagaacaa tattatagta

121
caacaaaata tgggtatgtc ataccctgcc acattcttga tgtaactttt tttcacctca

181
tgctcgccgg gttat.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a left sequence corresponding to SEQ ID NO: 14506 and a right sequence corresponding to SEQ ID NO: 14507. In certain embodiments, one piggyBac or piggyBac-like transposon end is at least 85%, at least 90%, at least 95%, at least 98%, at least 99% identical or any percentage in between identical to SEQ ID NO: 14506 and the other piggyBac or piggyBac-like transposon end is at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or any percentage in between identical to SEQ ID NO: 14507. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14506 and SEQ ID NO: 14507 or SEQ ID NO: 14509. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14508 and SEQ ID NO: 14507 or SEQ ID NO: 14509. In certain embodiments, the left and right transposon ends share a 16 bp repeat sequence at their ends of CCCGGCGAGCATGAGG (SEQ ID NO: 14510) immediately adjacent to the 5′-TTAT-3 target insertion site, which is inverted in the orientation in the two ends. In certain embodiments, left transposon end begins with a sequence comprising 5′-TTATCCCGGCGAGCATGAGG-3 (SEQ ID NO: 14511), and the right transposon ends with a sequence comprising the reverse complement of this sequence:

(SEQ ID NO: 14512)

5′-CCTCATGCTCGCCGGGTTAT-3′.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end comprising at least 14, 16, 18, 20, 30 or 40 contiguous nucleotides of SEQ ID NO: 14506 or SEQ ID NO: 14508. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end comprising at least 14, 16, 18, 20, 30 or 40 contiguous nucleotides of SEQ ID NO: 14507 or SEQ ID NO: 14509. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end with at least 90% identity to SEQ ID NO: 14506 or SEQ ID NO: 14508. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end with at least 90% identity to SEQ ID NO: 14507 or SEQ ID NO: 14509.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14515)

1
ttaacccggc gagcatgagg cagggtatct cataccctgg taaaatttta aagttgtgta

61
ttttataaaa ttttcgtctg acaacactag cgcgctcagt agctggaggc aggagcgtgc

121
gggaggggat agtggcgtga tcgcagtgtg gcacgggaca ccggcgagat attcgtgtgc

181
aaacctgttt cgggtatgtt ataccctgcc tcattgttga cgtatttttt ttatgtaatt

241
tttccgatta ttaatttcaa ctgttttatt ggtattttta tgttatccat tgttcttttt

301
ttatgattta ctgtatcggt tgtctttcgt tcctttagtt gagttttttt ttattatttt

361
cagtttttga tcaaa.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14516)

1
tcatattttt agtttaaaaa aataattata tgttttataa tgaaaagaat ctcattatct

61
ttcagtatta ggttgattta tattccaaag aataatattt ttgttaaatt gttgattttt

121
gtaaacctct aaatgtttgt tgctaaaatt actgtgttta agaaaaagat taataaataa

181
taataatttc ataattaaaa acttctttca ttgaatgcca ttaaataatt cattatttta

241
caaaataaga tcaacataat tgagtaaata ataataagaa caatattata gtacaacaaa

301
atatgggtat gtcataccct tttttttttt tttttttttt ttttttcggg tagagggccg

361
aacctcctac gaggtccccg cgcaaaaggg gcgcgcgggg tatgtgagac tcaacgatct

421
gcatggtgtt gtgagcagac cgcgggccca aggattttag agcccaccca ctaaacgact

481
cctctgcact cttacacccg acgtccgatc ccctccgagg tcagaacccg gatgaggtag

541
gggggctacc gcggtcaaca ctacaaccag acggcgcggc tcaccccaag gacgcccagc

601
cgacggagcc ttcgaggcga atcgaaggct ctgaaacgtc ggccgtctcg gtacggcagc

661
ccgtcgggcc gcccagacgg tgccgctggt gtcccggaat accccgctgg accagaacca

721
gcctgccggg tcgggacgcg atacaccgtc gaccggtcgc tctaatcact ccacggcagc

781
gcgctagagt gctggta.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of CCCGGCGAGCATGAGG (SEQ ID NO: 14510). In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of SEQ ID NO: 14510. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTATCCCGGCGAGCATGAGG (SEQ ID NO: 14511). In certain embodiments, the piggyBac or piggyBac-like transposon comprises at least 16 contiguous nucleotides from SEQ ID NO: 14511. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of CCTCATGCTCGCCGGGTTAT (SEQ ID NO: 14512). In certain embodiments, the piggyBac or piggyBac-like transposon comprises at least 16 contiguous nucleotides from SEQ ID NO: 14512. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end comprising at least 16 contiguous nucleotides from SEQ ID NO: 14511 and one end comprising at least 16 contiguous nucleotides from SEQ ID NO: 14512. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14511 and SEQ ID NO: 14512. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTAACCCGGCGAGCATGAGG (SEQ ID NO: 14513). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of CCTCATGCTCGCCGGGTTAA (SEQ ID NO: 14514).

In certain embodiments, the piggyBac or piggyBac-like transposon may have ends comprising SEQ ID NO: 14506 and SEQ ID NO: 14507, or a variant of either or both of these having at least 90% sequence identity to SEQ ID NO: 14506 or SEQ ID NO: 14507, and the piggyBac or piggyBac-like transposase has the sequence of SEQ ID NO: 14504 or SEQ ID NO: 14505, or a sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identity to SEQ ID NO: 14504 or SEQ ID NO: 14505. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a heterologous polynucleotide inserted between a pair of inverted repeats, where the transposon is capable of transposition by a piggyBac or piggyBac-like transposase having at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identity to SEQ ID NO: 14504 or SEQ ID NO: 14505. In certain embodiments, the transposon comprises two transposon ends, each of which comprises SEQ ID NO: 14510 in inverted orientations in the two transposon ends. In certain embodiments, each inverted terminal repeat (ITR) is at least 90% identical to SEQ ID NO: 14510.

In certain embodiments, the piggyBac or piggyBac-like transposon is capable of insertion by a piggyBac or piggyBac-like transposase at the sequence 5′-TTAT-3 within a target nucleic acid. In certain embodiments, one end of the piggyBac or piggyBac-like transposon comprises at least 16 contiguous nucleotides from SEQ ID NO: 14506 and the other transposon end comprises at least 16 contiguous nucleotides from SEQ ID NO: 14507. In certain embodiments, one end of the piggyBac or piggyBac-like transposon comprises at least 17, at least 18, at least 19, at least 20, at least 22, at least 25, at least 30 contiguous nucleotides from SEQ ID NO: 14506 and the other transposon end comprises at least 17, at least 18, at least 19, at least 20, at least 22, at least 25, at least 30 contiguous nucleotides from SEQ ID NO: 14507.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises transposon ends (each end comprising an ITR) corresponding to SEQ ID NO: 14506 and SEQ ID NO: 14507, and has a target sequence corresponding to 5′-TTAT3′. In certain embodiments, the piggyBac or piggyBac-like transposon also comprises a sequence encoding a transposase (e.g. SEQ ID NO: 14505). In certain embodiments, the piggyBac or piggyBac-like transposon comprises one transposon end corresponding to SEQ ID NO: 14506 and a second transposon end corresponding to SEQ ID NO: 14516. SEQ ID NO: 14516 is very similar to SEQ ID NO: 14507, but has a large insertion shortly before the ITR. Although the ITR sequences for the two transposon ends are identical (they are both identical to SEQ ID NO: 14510), they have different target sequences: the second transposon has a target sequence corresponding to 5′-TTAA-3′, providing evidence that no change in ITR sequence is necessary to modify the target sequence specificity. The piggyBac or piggyBac-like transposase (SEQ ID NO: 14504), which is associated with the 5′-TTAA-3′ target site differs from the 5′-TTAT-3′-associated transposase (SEQ ID NO: 14505) by only 4 amino acid changes (D322Y, S473C, A507T, H582R). In certain embodiments, the piggyBac or piggyBac-like transposase (SEQ ID NO: 14504), which is associated with the 5′-TTAA-3′ target site is less active than the 5′-TTAT-3′-associated piggyBac or piggyBac-like transposase (SEQ ID NO: 14505) on the transposon with 5′-TTAT-3′ ends. In certain embodiments, piggyBac or piggyBac-like transposons with 5′-TTAA-3′ target sites can be converted to piggyBac or piggyBac-like transposases with 5′-TTAT-3 target sites by replacing 5′-TTAA-3′ target sites with 5′-TTAT-3′. Such transposons can be used either with a piggyBac or piggyBac-like transposase such as SEQ ID NO: 14504 which recognizes the 5′-TTAT-3′ target sequence, or with a variant of a transposase originally associated with the 5′-TTAA-3′ transposon. In certain embodiments, the high similarity between the 5′-TTAA-3′ and 5′-TTAT-3′ piggyBac or piggyBac-like transposases demonstrates that very few changes to the amino acid sequence of a piggyBac or piggyBac-like transposase alter target sequence specificity. In certain embodiments, modification of any piggyBac or piggyBac-like transposon-transposase gene transfer system, in which 5′-TTAA-3′ target sequences are replaced with 5′-TTAT-3′-target sequences, the ITRs remain the same, and the transposase is the original piggyBac or piggyBac-like transposase or a variant thereof resulting from using a low-level mutagenesis to introduce mutations into the transposase. In certain embodiments, piggyBac or piggyBac-like transposon transposase transfer systems can be formed by the modification of a 5′-TTAT-3′-active piggyBac or piggyBac-like transposon-transposase gene transfer systems in which 5′-TTAT-3′ target sequences are replaced with 5′-TTAA-3′-target sequences, the ITRs remain the same, and the piggyBac or piggyBac-like transposase is the original transposase or a variant thereof.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Bombyx mori. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14577)

1
cccggcgagc atgaggcagg gtatctcata ccctggtaaa attttaaagt tgtgtatttt

61
ataaaatttt cgtctgacaa cactagcgcg ctcagtagct ggaggcagga gcgtgcggga

121
ggggatagtg gcgtgatcgc agtgtggcac gggacaccgg cgagatattc gtgtgcaaac

181
ctgtttcggg tatgttatac cctgcctcat tgttgacgta t.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14578)

1
tttaagaaaa agattaataa ataataataa tttcataatt aaaaacttct ttcattgaat

61
gccattaaat aaaccattat tttacaaaat aagatcaaca taattgagta aataataata

121
agaacaatat tatagtacaa caaaatatgg gtatgtcata ccctgccaca ttcttgatgt

181
aacttttttt cacctcatgc tcgccggg.

In certain embodiments, the transposon comprises at least 16 contiguous bases from SEQ ID NO: 14577 and at least 16 contiguous bases from SEQ ID NO: 14578, and inverted terminal repeats that are at least 87% identical to CCCGGCGAGCATGAGG (SEQ ID NO: 14510). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14595)

1
cccggcgagc atgaggcagg gtatctcata ccctggtaaa attttaaagt tgtgtatttt

61
ataaaatttt cgtctgacaa cactagcgcg ctcagtagct ggaggcagga gcgtgcggga

121
ggggatagtg gcgtgatcgc agtgtggcac gggacaccgg cgagatattc gtgtgcaaac

181
ctgtttcggg tatgttatac cctgcctcat tgttgacgta ttttttttat gtaatttttc

241
cgattattaa tttcaactgt tttattggta tttttatgtt atccattgtt ctttttttat

301
gatttactgt atcggttgtc tttcgttcct ttagttgagt ttttttttat tattttcagt

361
ttttgatcaa a.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14596)

1
tcatattttt agtttaaaaa aataattata tgttttataa tgaaaagaat ctcattatct

61
ttcagtatta ggttgattta tattccaaag aataatattt ttgttaaatt gttgattttt

121
gtaaacctct aaatgtttgt tgctaaaatt actgtgttta agaaaaagat taataaataa

181
taataatttc ataattaaaa acttctttca ttgaatgcca ttaaataaac cattatttta

241
caaaataaga tcaacataat tgagtaaata ataataagaa caatattata gtacaacaaa

301
atatgggtat gtcataccct gccacattct tgatgtaact ttttttcacc tcatgctcgc

361
cggg.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14595 and SEQ ID NO: 14596, and is transposed by the piggyBac or piggyBac-like transposase of SEQ ID NO: 14505. In certain embodiments, the ITRs of SEQ ID NO: 14595 and SEQ ID: 14596 are not flanked by a 5′-TTAA-3′ sequence. In certain embodiments, the ITRs of SEQ ID NO: 14595 and SEQ ID: 14596 are flanked by a 5′-TTAT-3′ sequence.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14597)

1
cccggcgagc atgaggcagg gtatctcata ccctggtaaa attttaaagt tgtgtatttt

61
ataaaatttt cgtctgacaa cactagcgcg ctcagtagct ggaggcagga gcgtgcggga

121
ggggatagtg gcgtgatcgc agtgtggcac gggacaccgg cgagatattc gtgtgcaaac

181
ctgtttcggg tatgttatac cctgcctcat tgttgacgta ttttttttat gtaatttttc

241
cgattattaa tttcaactgt tttattggta tttttatgtt atccattgtt ctttttttat

301
g.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14598)

1
cagggtatct cataccctgg taaaatttta aagttgtgta ttttataaaa ttttcgtctg

61
acaacactag cgcgctcagt agctggaggc aggagcgtgc gggaggggat agtggcgtga

121
tcgcagtgtg gcacgggaca ccggcgagat attcgtgtgc aaacctgttt cgggtatgtt

181
ataccctgcc tcattgttga cgtatttttt ttatgtaatt tttccgatta ttaatttcaa

241
ctgttttatt ggtattttta tgttatccat tgttcttttt ttatg.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14599)

1
cagggtatct cataccctgg taaaatttta aagttgtgta ttttataaaa ttttcgtctg

61
acaacactag cgcgctcagt agctggaggc aggagcgtgc gggaggggat agtggcgtga

121
tcgcagtgtg gcacgggaca ccggcgagat attcgtgtgc aaacctgttt cgggtatgtt

181
ataccctgcc tcattgttga cgtat.

In certain embodiments, the left end of the piggyBac or piggyBac-like transposon comprises a sequence of SEQ ID NO: 14577, SEQ ID NO: 14595, or SEQ ID NOs: 14597-14599. In certain embodiments, the left end of the piggyBac or piggyBac-like transposon is preceded by a left target sequence.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14600)

1
tcatattttt agtttaaaaa aataattata tgttttataa tgaaaagaat ctcattatct

61
ttcagtatta ggttgattta tattccaaag aataatattt ttgttaaatt gttgattttt

121
gtaaacctct aaatgtttgt tgctaaaatt actgtgttta agaaaaagat taataaataa

181
taataatttc ataattaaaa acttctttca ttgaatgcca ttaaataaac cattatttta

241
caaaataaga tcaacataat tgagtaaata ataataagaa caatattata gtacaacaaa

301
atatgggtat gtcataccct gccacattct tgatgtaact ttttttcacc tcatgctcgc

361
cggg.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14601)

1
tttaagaaaa agattaataa ataataataa tttcataatt aaaaacttct ttcattgaat

61
gccattaaat aaaccattat tttacaaaat aagatcaaca taattgagta aataataata

121
agaacaatat tatagtacaa caaaatatgg gtatgtcata ccctgccaca ttcttgatgt

181
aacttttttt ca.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14602)

1
cccggcgagc atgaggcagg gtatctcata ccctggtaaa attttaaagt tgtgtatttt

61
ataaaatttt cgtctgacaa cactagcgcg ctcagtagct ggaggcagga gcgtgcggga

121
ggggatagtg gcgtgatcgc agtgtggcac gggacaccgg cgagatattc gtgtgcaaac

181
ctgtttcggg tatgttatac cctgcctcat tgttgacgta ttttttttat gtaatttttc

241
cgattattaa tttcaactgt tttattggta tttttatgtt atccattgtt ctttttttat

301
gatttactgt atcggttgtc tttcgttcct ttagttgagt ttttttttat tattttcagt

361
ttttgatcaa a.

In certain embodiments, the right end of the piggyBac or piggyBac-like transposon comprises a sequence of SEQ ID NO: 14578, SEQ ID NO: 14596, or SEQ ID NOs: 14600-14601. In certain embodiments, the right end of the piggyBac or piggyBac-like transposon is followed by a right target sequence. In certain embodiments, the transposon is transposed by the transposase of SEQ ID NO: 14505. In certain embodiments, the left and right ends of the piggyBac or piggyBac-like transposon share a 16 bp repeat sequence of SEQ ID NO: 14510 in inverted orientation and immediately adjacent to the target sequence. In certain embodiments, the left transposon end begins with SEQ ID NO: 14510, and the right transposon end ends with the reverse complement of SEQ ID NO: 14510, 5′-CCTCATGCTCGCCGGG-3′ (SEQ ID NO: 14603). In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR with at least 93%, at least 87%, or at least 81% or any percentage in between identity to SEQ ID NO: 14510 or SEQ ID NO: 14603. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a target sequence followed by a left transposon end comprising a sequence selected from SEQ ID NOs: 88, 105 or 107 and a right transposon end comprising SEQ ID NO: 14578 or 106 followed by a target sequence. in certain embodiments, the piggyBac or piggyBac like transposon comprises one end that comprises a sequence that is at least 90%, at least 95% or at least 99% or any percentage in between identical to SEQ ID NO: 14577 and one end that comprises a sequence that is at least 90%, at least 95% or at least 99% or any percentage in between identical to SEQ ID NO: 14578. In certain embodiments, one transposon end comprises at least 14, at least 16, at least 18 or at least 20 contiguous bases from SEQ ID NO: 14577 and one transposon end comprises at least 14, at least 16, at least 18 or at least 20 contiguous bases from SEQ ID NO: 14578.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises two transposon ends wherein each transposon ends comprises a sequence that is at least 81% identical, at least 87% identical or at least 93% identical or any percentage in between identical to SEQ ID NO: 14510 in inverted orientation in the two transposon ends. One end may further comprise at least 14, at least 16, at least 18 or at least 20 contiguous bases from SEQ ID NO: 14599, and the other end may further comprise at least 14, at least 16, at least 18 or at least 20 contiguous bases from SEQ ID NO: 14601. The piggyBac or piggyBac-like transposon may be transposed by the transposase of SEQ ID NO: 14505, and the transposase may optionally be fused to a nuclear localization signal.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14595 and SEQ ID NO: 14596 and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14504 or SEQ ID NO: 14505. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14597 and SEQ ID NO: 14596 and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14504 or SEQ ID NO: 14505. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14595 and SEQ ID NO: 14578 and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14504 or SEQ ID NO: 14505. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14602 and SEQ ID NO: 14600 and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14504 or SEQ ID NO: 14505.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a left end comprising 1, 2, 3, 4, 5, 6, or 7 sequences selected from ATGAGGCAGGGTAT (SEQ ID NO: 14614), ATACCCTGCCTCAT (SEQ ID NO: 14615), GGCAGGGTAT (SEQ ID NO: 14616), ATACCCTGCC (SEQ ID NO: 14617), TAAAATTTTA (SEQ ID NO: 14618), ATTTTATAAAAT (SEQ ID NO: 14619), TCATACCCTG (SEQ ID NO: 14620) and TAAATAATAATAA (SEQ ID NO: 14621). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a right end comprising 1, 2 or 3 sequences selected from SEQ ID NO: 14617, SEQ ID NO: 14620 and SEQ ID NO: 14621.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Xenopus tropicalis. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14517)

1
MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQNPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SLESYWDTTT VLSIPVFSAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241
SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHY.

In some embodiments, the piggyBac or piggyBac-like transposase is a hyperactive variant of SEQ ID NO: 14517. In certain embodiments, the piggyBac or piggyBac-like transposase is an integration defective variant of SEQ ID NO: 14517. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14518)

1
MAKRFYSAEE AAAHCMAPSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQNPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SLESYWNTTT VLSIPVFSAT MSRNRYQLLL RFLHFNNNAT AVPPDQPDHD RLHKLRPLID

241
SLSERFAAVY TPCQNICIDE SLLLFKGRLR FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT

361
PACGTINRTR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT SAWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMLP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHY.

In certain embodiments, the piggyBac or piggyBac-like transposase is isolated or derived from Xenopus tropicalis. In certain embodiments, the piggyBac or piggyBac-like transposase is a hyperactive piggyBac or piggyBac-like transposase. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence at least 90% identical to:

(SEQ ID NO: 14572)

1
MAKRFYSAEE AAAHCSASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQNPLTRG ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SIESYWDTTT VLSIPVFGAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241
SLSERFANVY TPCQNICIDE SLMLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSTGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPD SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCRKPCF EIYHTQLHY.

In certain embodiments, piggyBac or piggyBac-like transposase is a hyperactive piggyBac or piggyBac-like transposase. A hyperactive piggyBac or piggyBac-like transposase is a transposase that is more active than the naturally occurring variant from which it is derived. In certain embodiments, a hyperactive piggyBac or piggyBac-like transposase is more active than the transposase of SEQ ID NO: 14517. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 14624)

1
MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQNPLTRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SLESYWDTTT VLSIPVFSAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241
SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCRKPCF EIYHTQLHY.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 14625)

1
MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQNPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SLESYWDTTT VLKIPVFSAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241
SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHY.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 14627)

1
MAKRFYSAEE AAAHCMASSS EQTSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQNPLTRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SIESYWDTTT VLSIPVFGAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241
SLSERFANVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRKPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCRKPCF EIYHTQLHY.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 14628)

1
MAKRFYSAEE AAAHCSASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQNPLTRG ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SLESYWDTTT VLSIPVFGAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241
SLSERFANVY TPCQNICIDE SLMLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSTGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCRKPCF EIYHTQLHY.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

(SEQ ID NO: 149)

1
MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQNPLTRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SLESYWDTTT VLSIPVFGAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241
SLSERFANVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCRKPCF EIYHTQLHY.

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises an amino acid substitution at a position selected from amino acid 6, 7, 16, 19, 20, 21, 22, 23, 24, 26, 28, 31, 34, 67, 73, 76, 77, 88, 91, 141, 145, 146, 148, 150, 157, 162, 179, 182, 189, 192, 193, 196, 198, 200, 210, 212, 218, 248, 263, 270, 294, 297, 308, 310, 333, 336, 354, 357, 358, 359, 377, 423, 426, 428, 438, 447, 450, 462, 469, 472, 498, 502, 517, 520, 523, 533, 534, 576, 577, 582, 583 or 587 (relative to SEQ ID NO: 14517). In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises an amino acid substitution of Y6C, S7G, M165, S19G, 520Q, 520G, 520D, E21D, E22Q, F23T, F23P, S24Y, S26V, S28Q, V31K, A34E, L67A, G73H, A76V, D77N, P88A, N91D, Y141Q, Y141A, N145E, N145V, P146T, P146V, P146K, P148T, P148H, Y150G, Y1505, Y150C, H157Y, A162C, A179K, L182I, L182V, T189G, L192H, S193N, S193K, V196I, S198G, T200W, L210H, F212N, N218E, A248N, L263M, Q270L, S294T, T297M, 5308R, L310R, L333M, Q336M, A354H, C357V, L358F, D359N, L377I, V423H, P426K, K428R, S438A, T447G, T447A, L450V, A462H, A462Q, I469V, I472L, Q498M, L502V, E5171, P520D, P520G, N523S, 1533E, D534A, F576R, F576E, K5771, I582R, Y583F, L587Y or L587W, or any combination thereof including at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or all of these mutations (relative to SEQ ID NO: 14517).

In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises one or more substitutions of an amino acid that is not wild type, wherein the one or more substitutions a for wild type amino acid comprises a substitution of A2X, K3X, R4X, FSX, Y6X, S7X, A11X, A13X, C15X, M16X, A17X, 518X, 519X, 520X, E21X, E22X, F23X, S24X, G25X, 26X, D27X, S28X, E29X, E42X, E43X, S44X, C46X, S47X, S48X, S49X, T50X, V51X, S52X, A53X, L54X, E55X, E56X, P57X, M58X, E59X, E62X, D63X, V64X, D65X, D66X, L67X, E68X, D69X, Q70X, E71X, A72X, G73X, D74X, R75X, A76X, D77X, A78X, A79X, A80X, G81X, G82X, E83X, P84X, A85X, W86X, G87X, P88X, P89X, C90X, N91X, F92X, P93X, E95X, I96X, P97X, P98X, F99X, T100X, T101X, P103X, G104X, V105X, K106X, V107X, D108X, T109X, N111X, P114X, Il 15X, N116X, F117X, F118X, Q119X, M122X, T123X, E124X, A125X, I126X, L127X, Q128X, D129X, M130X, L132X, Y133X, V126X, Y127X, A138X, E139X, Q140X, Y141X, L142X, Q144X, N145X, P146X, L147X, P148X, Y150X, A151X, A155X, H157X, P158X, I161X, A162X, V168X, T171X, L172X, A173X, M174X, I177X, A179X, L182X, D187X, T188X, T189X, T190X, L192X, S193X, I194X, P195X, V196X, S198X, A199X, T200X, S202X, L208X, L209X, L210X, R211X, F212X, F215X, N217X, N218X, A219X, T220X, A221X, V222X, P224X, D225X, Q226X, P227X, H229X, R231X, H233X, L235X, P237X, I239X, D240X, L242X, S243X, E244X, R244X, F246X, A247X, A248X, V249X, Y250X, T251X, P252X, C253X, Q254X, I256X, C257X, I258X, D259X, E260X, S261X, L262X, L263X, L264X, F265X, K266X, G267X, R268X, L269X, Q270X, F271X, R272X, Q273X, Y274X, I275X, P276X, S277X, K278X, R279X, A280X, R281X, Y282X, G283X, I284X, K285X, F286X, Y287X, K288X, L289X, C290X, E291X, S292X, S293XS294X, G295X, Y296X, T297X, S298X, Y299X, F300X, E304X, L310X, P313X, G314X, P316X, P317X, D318X, L319X, T320X, V321X, K324X, E328X, I330X, S331X, P332X, L333X, L334X, G335X, Q336X, F338X, L340X, D343X, N344X, F345X, Y346X, S347X, L351X, F352X, A354X, L355X, Y356X, C357X, L358X, D359X, T360X, R422X, Y423X, G424X, P426X, K428X, N429X, K430X, P431X, L432X, S434X, K435X, E436X, S438X, K439X, Y440X, G443X, R446X, T447X, L450X, Q451X, N455X, T460X, R461X, A462X, K465X, V467X, G468X, I469X, Y470X, L471X, I472X, M474X, A475X, L476X, R477X, S479X, Y480X, V482XY483X, K484X, A485X, A486X, V487X, P488X, P490X, K491X, S493X, Y494X, Y495X, K496X, Y497T, Q498X, L499X, Q500X, I501X, L502X, P503X, A504X, L505X, L506X, F507X, G508X, G509X, V510X, E511X, E512X, Q513X, T514X, V515X, E517X, M518X, P519X, P520X, S521X, D522X, N523X, V524X, A525X, L527X, I528X, K530X, H531X, F532X, I533X, D534X, T535X, L536X, T539X, P540X, Q546X, K550X, R553X, K554X, R555X, G556X, I557X, R558X, R559X, D560X, T561X, Y564X, P566X, K567X, P569X, R570X, N571X, L574X, C575X, F576X, K577X, P578X, F580X, E581X, I582X, Y583X, T585X, Q586X, L587X, H588X or Y589X (relative to SEQ ID NO: 14517). A list of hyperactive amino acid substitutions can be found in U.S. Pat. No. 10,041,077, the contents of which are incorporated by reference in their entirety.

In certain embodiments, the piggyBac or piggyBac-like transposase is integration deficient. In certain embodiments, an integration deficient piggyBac or piggyBac-like transposase is a transposase that can excise its corresponding transposon, but that integrates the excised transposon at a lower frequency than a corresponding naturally occurring transposase. In certain embodiments, the piggyBac or piggyBac-like transposase is an integration deficient variant of SEQ ID NO: 14517. In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase is deficient relative to SEQ ID NO: 14517.

In certain embodiments, the piggyBac or piggyBac-like transposase is active for excision but deficient in integration. In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to a sequence of:

(SEQ ID NO: 14605)

1
MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRVDAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQNPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SLESYWDTTT VLSIPVFSAT MSRNRYQLLL KFLHFNNEAT AVPPDQPGHD RLHKLRPLID

241
SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHYG RR.

In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to a sequence of:

(SEQ ID NO: 14604)

1
MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQVPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SLESYWDTTT VLNIPVFSAT MSRNRYQLLL RFLEFNNEAT AVPPDQPGHD RLHKLRPLID

241
SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHY.

In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to a sequence of:

(SEQ ID NO: 14611)

1
MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQNVLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SLESYWDTTT VLSIPVFSAT MSRNRYQLLL RFLHFNNDAT AVPPDQPGHD RLHKLRPLID

241
SLTERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHYG RR.

In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14611. In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to a sequence of:

(SEQ ID NO: 14612)

1
MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAP GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQVPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SLESYWDTTT VLSIPVFSAT MSRNRYQLLL RFLHFNNEAT AVPPDQPGHD RLHKLRPLID

241
SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHYG RR.

In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14612. In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to a sequence of:

(SEQ ID NO: 14613)

1
MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61
DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121
FMTEAILQDM VLYTNVYAEQ YLTQVPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181
SLESYWDTTT VLNIPVFSAT MSRNRYQLLL RFLEFNNNAT AVPPDQPGHD RLHKLRPLID

241
SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301
LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT

361
PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421
QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY

481
IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541
GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHYG RR.

In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14613. In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises an amino acid substitution wherein the Asn at position 218 is replaced by a Glu or an Asp (N218D or N218E) (relative to SEQ ID NO: 14517).

In certain embodiments, the excision competent, integration deficient piggyBac or piggyBac-like transposase comprises one or more substitutions of an amino acid that is not wild type, wherein the one or more substitutions a for wild type amino acid comprises a substitution of A2X, K3X, R4X, FSX, Y6X, S7X, ABX, E9X, E10X, A11X, A12X, A13X, H14X, C15X, M16X, A17X, 518X, 519X, 520X, E21X, E22X, F23X, S24X, G25X, 26X, D27X, S28X, E29X, V31X, P32X, P33X, A34X, 535X, E36X, S37X, D38X, S39X, 540X, T41X, E42X, E43X, S44X, W45X, C46X, S47X, S48X, S49X, T50X, V51X, S52X, A53X, L54X, E55X, E56X, P57X, M58X, E59X, V60X, M122X, T123X, E124X, A125X, L127X, Q128X, D129X, L132X, Y133X, V126X, Y127X, E139X, Q140X, Y141X, L142X, T143X, Q144X, N145X, P146X, L147X, P148X, R149X, Y150X, A151X, H154X, H157X, P158X, T159X, D160X, I161X, A162X, E163X, M164X, K165X, R166X, F167X, V168X, G169X, L170X, T171X, L172X, A173X, M174X, G175X, L176X, I177X, K178X, A179X, N180X, 5181X, L182X, 5184X, Y185X, D187X, T188X, T189X, T190X, V191X, L192X, 5193X, I194X, P195X, V196X, F197X, 5198X, A199X, T200X, M201X, 5202X, R203X, N204X, R205X, Y206X, Q207X, L208X, L209X, L210X, R211X, F212X, L213X, H241X, F215X, N216X, N217X, N218X, A219X, T220X, A221X, V222X, P223X, P224X, D225X, Q226X, P227X, G228X, H229X, D230X, R231X, H233X, K234X, L235X, R236X, L238X, I239X, D240X, L242X, S243X, E244X, R244X, F246X, A247X, A248X, V249X, Y250X, T251X, P252X, C253X, Q254X, N255X, I256X, C257X, I258X, D259X, E260X, S261X, L262X, L263X, L264X, F265X, K266X, G267X, R268X, L269X, Q270X, F271X, R272X, Q273X, Y274X, I275X, P276X, S277X, K278X, R279X, A280X, R281X, Y282X, G283X, I284X, K285X, F286X, Y287X, K288X, L289X, C290X, E291X, S292X, S293X, S294X, G295X, Y296X, T297X, S298X, Y299X, F300X, I302X, E304X, G305X, K306X, D307X, S308X, K309X, L310X, D311X, P312X, P313X, G314X, C315X, P316X, P317X, D318X, L319X, T320X, V321X, S322X, G323X, K324X, I325X, V326X, W327X, E328X, L329X, 1330X, S331X, P332X, L333X, L334X, G335X, Q336X, F338X, H339X, L340X, V342X, N344X, F345X, Y346X, S347X, S348X, I349X, L351X, T353X, A354X, Y356X, C357X, L358X, D359X, T360X, P361X, A362X, C363X, G364X, I366X, N367X, R368X, D369X, K371X, G372X, L373X, R375X, A376X, L377X, L378X, D379X, K380X, K381X, L382X, N383X, R384XG385X, T387X, Y388X, A389X, L390X, K392X, N393X, E394X, A397X, K399X, F400X, F401X, D402X, N405X, L406X, L409X, R422X, Y423X, G424X, E425X, P426X, K428X, N429X, K430X, P431X, L432X, S434X, K435X, E436X, S438X, K439X, Y440X, G442X, G443X, V444X, R446X, T447X, L450X, Q451X, H452X, N455X, T457X, R458X, T460X, R461X, A462X, Y464X, K465X, V467X, G468X, I469X, L471X, I472X, Q473X, M474X, L476X, R477X, N478X, S479X, Y480X, V482XY483X, K484X, A485X, A486X, V487X, P488X, G489X, P490X, K491X, L492X, S493X, Y494X, Y495X, K496X, Q498X, L499X, Q500X, I501X, L502X, P503X, A504X, L505X, L506X, F507X, G508X, G509X, V510X, E511X, E512X, Q513X, T514X, V515X, E517X, M518X, P519X, P520X, S521X, D522X, N523X, V524X, A525X, L527X, I528X, G529X, K530X, F532X, I533X, D534X, T535X, L536X, P537X, P538X, T539X, P540X, G541X, F542X, Q543X, R544X, P545X, Q546X, K547X, G548X, C549X, K550X, V551X, C552X, R553X, K554X, R555X, G556X, I557X, R558X, R559X, D560X, T561X, R562X, Y563X, Y564X, C565X, P566X, K567X, C568X, P569X, R570X, N571X, P572X, G573X, L574X, C575X, F576X, K577X, P578X, C579X, F580X, E581X, I582X, Y583X, H584X, T585X, Q586X, L587X, H588X or Y589X (relative to SEQ ID NO: 14517). A list of excision competent, integration deficient amino acid substitutions can be found in U.S. Pat. No. 10,041,077, the contents of which are incorporated by reference in their entirety.

In certain embodiments, the piggyBac or piggyBac-like transposase is fused to a nuclear localization signal. In certain embodiments, SEQ ID NO: 14517 or SEQ ID NO: 14518 is fused to a nuclear localization signal. In certain embodiments, the amino acid sequence of the piggyBac or piggyBac like transposase fused to a nuclear localization signal is encoded by a polynucleotide sequence comprising:

(SEQ ID NO: 14626)

1
atggcaccca aaaagaaacg taaagtgatg gccaaaagat tttacagcgc cgaagaagca

61
gcagcacatt gcatggcatc gtcatccgaa gaattctcgg ggagcgattc cgaatatgtc

121
ccaccggcct cggaaagcga ttcgagcact gaggagtcgt ggtgttcctc ctcaactgtc

181
tcggctcttg aggagccgat ggaagtggat gaggatgtgg acgacttgga ggaccaggaa

241
gccggagaca gggccgacgc tgccgcggga ggggagccgg cgtggggacc tccatgcaat

301
tttcctcccg aaatcccacc gttcactact gtgccgggag tgaaggtcga cacgtccaac

361
ttcgaaccga tcaatttctt tcaactcttc atgactgaag cgatcctgca agatatggtg

421
ctctacacta atgtgtacgc cgagcagtac ctgactcaaa acccgctgcc tcgctacgcg

481
agagcgcatg cgtggcaccc gaccgatatc gcggagatga agcggttcgt gggactgacc

541
ctcgcaatgg gcctgatcaa ggccaacagc ctcgagtcat actgggatac cacgactgtg

601
cttagcattc cggtgttctc cgctaccatg tcccgtaacc gctaccaact cctgctgcgg

661
ttcctccact tcaacaacaa tgcgaccgct gtgccacctg accagccagg acacgacaga

721
ctccacaagc tgcggccatt gatcgactcg ctgagcgagc gattcgccgc ggtgtacacc

781
ccttgccaaa acatttgcat cgacgagtcg cttctgctgt ttaaaggccg gcttcagttc

841
cgccagtaca tcccatcgaa gcgcgctcgc tatggtatca aattctacaa actctgcgag

901
tcgtccagcg gctacacgtc atacttcttg atctacgagg ggaaggactc taagctggac

961
ccaccggggt gtccaccgga tcttactgtc tccggaaaaa tcgtgtggga actcatctca

1021
cctctcctcg gacaaggctt tcatctctac gtcgacaatt tctactcatc gatccctctg

1081
ttcaccgccc tctactgcct ggatactcca gcctgtggga ccattaacag aaaccggaag

1141
ggtctgccga gagcactgct ggataagaag ttgaacaggg gagagactta cgcgctgaga

1201
aagaacgaac tcctcgccat caaattcttc gacaagaaaa atgtgtttat gctcacctcc

1261
atccacgacg aatccgtcat ccgggagcag cgcgtgggca ggccgccgaa aaacaagccg

1321
ctgtgctcta aggaatactc caagtacatg gggggtgtcg accggaccga tcagctgcag

1381
cattactaca acgccactag aaagacccgg gcctggtaca agaaagtcgg catctacctg

1441
atccaaatgg cactgaggaa ttcgtatatt gtctacaagg ctgccgttcc gggcccgaaa

1501
ctgtcatact acaagtacca gcttcaaatc ctgccggcgc tgctgttcgg tggagtggaa

1561
gaacagactg tgcccgagat gccgccatcc gacaacgtgg cccggttgat cggaaagcac

1621
ttcattgata ccctgcctcc gacgcctgga aagcagcggc cacagaaggg atgcaaagtt

1681
tgccgcaagc gcggaatacg gcgcgatacc cgctactatt gcccgaagtg cccccgcaat

1741
cccggactgt gtttcaagcc ctgttttgaa atctaccaca cccagttgca ttac.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Xenopus tropicalis. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14519)

1
ttaacctttt tactgccaat gacgcatggg atacgtcgtg gcagtaaaag ggcttaaatg

61
ccaacgacgc gtcccatacg ttgttggcat tttaagtctt ctctctgcag cggcagcatg

121
tgccgccgct gcagagagtt tctagcgatg acagcccctc tgggcaacga gccggggggg

181
ctgtc.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14520)

1
tttgcatttt tagacattta gaagcctata tcttgttaca gaattggaat tacacaaaaa

61
ttctaccata ttttgaaagc ttaggttgtt ctgaaaaaaa caatatattg ttttcctggg

121
taaactaaaa gtcccctcga ggaaaggccc ctaaagtgaa acagtgcaaa acgttcaaaa

181
actgtctggc aatacaagtt ccactttgac caaaacggct ggcagtaaaa gggttaa.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14519 and SEQ ID NO: 14520. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14521)

1
ttaacccttt gcctgccaat cacgcatggg atacgtcgtg gcagtaaaag ggcttaaatg

61
ccaacgacgc gtcccatacg ttgttggcat tttaagtctt ctctctgcag cggcagcatg

121
tgccgccgct gcagagagtt tctagcgatg acagcccctc tgggcaacga gccggggggg

181
ctgtc.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14522)

1
tttgcatttt tagacattta gaagcctata tcttgttaca gaattggaat tacacaaaaa

61
ttctaccata ttttgaaagc ttaggttgtt ctgaaaaaaa caatatattg ttttcctggg

121
taaactaaaa gtcccctcga ggaaaggccc ctaaagtgaa acagtgcaaa acgttcaaaa

181
actgtctggc aatacaagtt ccactttggg acaaatcggc tggcagtgaa agggttaa.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14523)

1
ttaacctttt tactgccaat gacgcatggg atacgtcgtg gcagtaaaag ggcttaaatg

61
ccaacgacgc gtcccatacg ttgttggcat tttaattctt ctctctgcag cggcagcatg

121
tgccgccgct gcagagagtt tctagcgatg acagcccctc tgggcaacga gccggggggg

181
ctgtc.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14520 and SEQ ID NO: 14519, SEQ ID NO: 14521 or SEQ ID NO: 14523. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14522 and SEQ ID NO: 14519, SEQ ID NO: 14521 or SEQ ID NO: 14523. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end comprising at least 14, 16, 18, 20, 30 or 40 contiguous nucleotides from SEQ ID NO: 14519, SEQ ID NO: 14521 or SEQ ID NO: 14523. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end comprising at least 14, 16, 18, 20, 30 or 40 contiguous nucleotides from SEQ ID NO: 14520 or SEQ ID NO: 14522. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end with at least 90% identity to SEQ ID NO: 14519, SEQ ID NO: 14521 or SEQ ID NO: 14523. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end with at least 90% identity to SEQ ID NO: 14520 or SEQ ID NO: 14522. In one embodiment, one transposon end is at least 90% identical to SEQ ID NO: 14519 and the other transposon end is at least 90% identical to SEQ ID NO: 14520.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTAACCTTTTTACTGCCA (SEQ ID NO: 14524). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTAACCCTTTGCCTGCCA (SEQ ID NO: 14526). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTAACCYTTTTACTGCCA (SEQ ID NO: 14527). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TGGCAGTAAAAGGGTTAA (SEQ ID NO: 14529). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TGGCAGTGAAAGGGTTAA (SEQ ID NO: 14531). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTAACCYTTTKMCTGCCA (SEQ ID NO: 14533). In certain embodiments, one end of the piggyBac or piggyBac-like transposon comprises a sequence selected from SEQ ID NO: 14524, SEQ ID NO: 14526 and SEQ ID NO: 14527. In certain embodiments, one end of the piggyBac™ (PB) or piggyBac-like transposon comprises a sequence selected from SEQ ID NO: 14529 and SEQ ID NO: 14531. In certain embodiments, each inverted terminal repeat of the piggyBac or piggyBac-like transposon comprises a sequence of ITR sequence of CCYTTTKMCTGCCA (SEQ ID NO: 14563). In certain embodiments, each end of the piggyBac™ (PB) or piggyBac-like transposon comprises SEQ ID NO: 14563 in inverted orientations. In certain embodiments, one ITR of the piggyBac or piggyBac-like transposon comprises a sequence selected from SEQ ID NO: 14524, SEQ ID NO: 14526 and SEQ ID NO: 14527. In certain embodiments, one ITR of the piggyBac or piggyBac-like transposon comprises a sequence selected from SEQ ID NO: 14529 and SEQ ID NO: 14531. In certain embodiments, the piggyBac or piggyBac like transposon comprises SEQ ID NO: 14533 in inverted orientation in the two transposon ends.

In certain embodiments, The piggyBac or piggyBac-like transposon may have ends comprising SEQ ID NO: 14519 and SEQ ID NO: 14520 or a variant of either or both of these having at least 90% sequence identity to SEQ ID NO: 14519 or SEQ ID NO: 14520, and the piggyBac or piggyBac-like transposase has the sequence of SEQ ID NO: 14517 or a variant showing at least %, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between sequence identity to SEQ ID NO: 14517 or SEQ ID NO: 14518. In certain embodiments, one piggyBac or piggyBac-like transposon end comprises at least 14 contiguous nucleotides from SEQ ID NO: 14519, SEQ ID NO: 14521 or SEQ ID NO: 14523, and the other transposon end comprises at least 14 contiguous nucleotides from SEQ ID NO: 14520 or SEQ ID NO: 14522. In certain embodiments, one transposon end comprises at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 22, at least 25, at least 30 contiguous nucleotides from SEQ ID NO: 14519, SEQ ID NO: 14521 or SEQ ID NO: 14523, and the other transposon end comprises at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 22, at least 25 or at least 30 contiguous nucleotides from SEQ ID NO: 14520 or SEQ ID NO: 14522.

In certain embodiments, the piggyBac or piggyBac-like transposase recognizes a transposon end with a left sequence corresponding to SEQ ID NO: 14519, and a right sequence corresponding to SEQ ID NO: 14520. It will excise the transposon from one DNA molecule by cutting the DNA at the 5′-TTAA-3′ sequence at the left end of one transposon end to the 5′-TTAA-3′ at the right end of the second transposon end, including any heterologous DNA that is placed between them, and insert the excised sequence into a second DNA molecule. In certain embodiments, truncated and modified versions of the left and right transposon ends will also function as part of a transposon that can be transposed by the piggyBac or piggyBac-like transposase. For example, the left transposon end can be replaced by a sequence corresponding to SEQ ID NO: 14521 or SEQ ID NO: 14523, the right transposon end can be replaced by a shorter sequence corresponding to SEQ ID NO: 14522. In certain embodiments, the left and right transposon ends share an 18 bp almost perfectly repeated sequence at their ends (5′-TTAACCYTTTKMCTGCCA: SEQ ID NO: 14533) that includes the 5′-TTAA-3′ insertion site, which sequence is inverted in the orientation in the two ends. That is in SEQ ID NO: 14519 and SEQ ID NO: 14523 the left transposon end begins with the sequence 5′-TTAACCTTTTTACTGCCA-3′ (SEQ ID NO: 14524), or in SEQ ID NO: 14521 the left transposon end begins with the sequence 5′-TTAACCCTTTGCCTGCCA-3′ (SEQ ID NO: 14526); the right transposon ends with approximately the reverse complement of this sequence: in SEQ ID NO: 14520 it ends 5′ TGGCAGTAAAAGGGTTAA-3′ (SEQ ID NO: 14529), in SEQ ID NO: 14522 it ends 5′-TGGCAGTGAAAGGGTTAA-3′ (SEQ ID NO: 14531.) One embodiment of the invention is a transposon that comprises a heterologous polynucleotide inserted between two transposon ends each comprising SEQ ID NO: 14533 in inverted orientations in the two transposon ends. In certain embodiments, one transposon end comprises a sequence selected from SEQ ID NOS: 14524, SEQ ID NO: 14526 and SEQ ID NO: 14527. In some embodiments, one transposon end comprises a sequence selected from SEQ ID NO: 14529 and SEQ ID NO: 14531.

In certain embodiments, the piggyBac™ (PB) or piggyBac-like transposon is isolated or derived from Xenopus tropicalis. In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14573)

1
ccctttgcct gccaatcacg catgggatac gtcgtggcag taaaagggct taaatgccaa

61
cgacgcgtcc catacgtt.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14574)

1
cctgggtaaa ctaaaagtcc cctcgaggaa aggcccctaa agtgaaacag tgcaaaacgt

61
tcaaaaactg tctggcaata caagttccac tttgggacaa atcggctggc agtgaaaggg.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at least 16 contiguous bases from SEQ ID NO: 14573 or SEQ ID NO: 14574, and inverted terminal repeat of CCYTTTBMCTGCCA (SEQ ID NO: 14575).

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14579)

1
ccctttgcct gccaatcacg catgggatac gtcgtggcag taaaagggct taaatgccaa

61
cgacgcgtcc catacgttgt tggcatttta agtcttctct ctgcagcggc agcatgtgcc

121
gccgctgcag agagtttcta gcgatgacag cccctctggg caacgagccg ggggggctgt

181
c.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14580)

1
cctttttact gccaatgacg catgggatac gtcgtggcag taaaagggct taaatgccaa

61
cgacgcgtcc catacgttgt tggcatttta attcttctct ctgcagcggc agcatgtgcc

121
gccgctgcag agagtttcta gcgatgacag cccctctggg caacgagccg ggggggctgt

181
c.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14581)

1
cctttttact gccaatgacg catgggatac gtcgtggcag taaaagggct taaatgccaa

61
cgacgcgtcc catacgttgt tggcatttta agtcttctct ctgcagcggc agcatgtgcc

121
gccgctgcag agagtttcta gcgatgacag cccctctggg caacgagccg ggggggctgt

181
c.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14582)

1
cctttttact gccaatgacg catgggatac gtcgtggcag taaaagggct taaatgccaa

61
cgacgcgtcc catacgttgt tggcatttta agtcttctct ctgcagcggc agcatgtgcc

121
gccgctgcag agag.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14583)

1
cctttttact gccaatgacg catgggatac gtcgtggcag taaaagggct taaatgccaa

61
cgacgcgtcc catacgttgt tggcatttta agtctt.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14584)

1
ccctttgcct gccaatcacg catgggatac gtcgtggcag taaaagggct taaatgccaa

61
cgacgcgtcc catacgttgt tggcatttta agtctt .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14585)

1
ttatcctttt tactgccaat gacgcatggg atacgtcgtg gcagtaaaag ggcttaaatg

61
ccaacgacgc gtcccatacg ttgttggcat tttaagtctt ctctctgcag cggcagcatg

121
tgccgccgct gcagagagtt tctagcgatg acagcccctc tgggcaacga gccggggggg

181
ctgtc.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14586)

1
tttgcatttt tagacattta gaagcctata tcttgttaca gaattggaat tacacaaaaa

61
ttctaccata ttttgaaagc ttaggttgtt ctgaaaaaaa caatatattg ttttcctggg

121
taaactaaaa gtcccctcga ggaaaggccc ctaaagtgaa acagtgcaaa acgttcaaaa

181
actgtctggc aatacaagtt ccactttggg acaaatcggc tggcagtgaa aggg.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a left transposon end sequence selected from SEQ ID NO: 14573 and SEQ ID NOs: 14579-14585. In certain embodiments, the left transposon end sequence is preceded by a left target sequence. In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14587)

1
tttgcatttt tagacattta gaagcctata tcttgttaca gaattggaat tacacaaaaa

61
ttctaccata ttttgaaagc ttaggttgtt ctgaaaaaaa caatatattg ttttcctggg

121
taaactaaaa gtcccctcga ggaaaggccc ctaaagtgaa acagtgcaaa acgttcaaaa

181
actgtctggc aatacaagtt ccactttgac caaaacggct ggcagtaaaa ggg.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14588)

1
ttgttctgaa aaaaacaata tattgttttc ctgggtaaac taaaagtccc ctcgaggaaa

61
ggcccctaaa gtgaaacagt gcaaaacgtt caaaaactgt ctggcaatac aagttccact

121
ttgaccaaaa cggctggcag taaaaggg.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14589)

1
tttgcatttt tagacattta gaagcctata tcttgttaca gaattggaat tacacaaaaa

61
ttctaccata ttttgaaagc ttaggttgtt ctgaaaaaaa caatatattg ttttcctggg

121
taaactaaaa gtcccctcga ggaaaggccc ctaaagtgaa acagtgcaaa acgttcaaaa

181
actgtctggc aatacaagtt ccactttgac caaaacggct ggcagtaaaa gggttat.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

(SEQ ID NO: 14590)

1
ttgttctgaa aaaaacaata tattgttttc ctgggtaaac taaaagtccc ctcgaggaaa

61
ggcccctaaa gtgaaacagt gcaaaacgtt caaaaactgt ctggcaatac aagttccact

121
ttgggacaaa tcggctggca gtgaaaggg.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a right transposon end sequence selected from SEQ ID NO: 14574 and SEQ ID NOs: 14587-14590. In certain embodiments, the right transposon end sequence is followed by a right target sequence. In certain embodiments, the left and right transposon ends share a 14 repeated sequence inverted in orientation in the two ends (SEQ ID NO: 14575) adjacent to the target sequence. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a left transposon end comprising a target sequence and a sequence that is selected from SEQ ID NOs: 14582-14584 and 14573, and a right transposon end comprising a sequence selected from SEQ ID NOs: 14588-14590 and 14574 followed by a right target sequence.

In certain embodiments, the left transposon end of the piggyBac or piggyBac-like transposon comprises

1
atcacgcatg ggatacgtcg tggcagtaaa agggcttaaa tgccaacgac gcgtcccata

61
cgtt

(SEQ ID NO: 14591), and an ITR. In certain embodiments, the left transposon end comprises

1
atgacgcatg ggatacgtcg tggcagtaaa agggcttaaa tgccaacgac gcgtcccata

61
cgttgttggc attttaagtc tt

(SEQ ID NO: 14592) and an ITR. In certain embodiments, the right transposon end of the piggyBac or piggyBac-like transposon comprises

1
cctgggtaaa ctaaaagtcc cctcgaggaa aggcccctaa agtgaaacag tgcaaaacgt

61
tcaaaaactg tctggcaata caagttccac tttgggacaa atcggc

(SEQ ID NO: 14593) and an ITR. In certain embodiments, the right transposon end comprises

1
ttgttctgaa aaaaacaata tattgttttc ctgggtaaac taaaagtccc ctcgaggaaa

61
ggcccctaaa gtgaaacagt gcaaaacgtt caaaaactgt ctggcaatac aagttccact

121
ttgaccaaaa cggc

(SEQ ID NO: 14594) and an ITR.

In certain embodiments, one transposon end comprises a sequence that is at least 90%, at least 95%, at least 99% or any percentage in between identical to SEQ ID NO: 14573 and the other transposon end comprises a sequence that is at least 90%, at least 95%, at least 99% or any percentage in between identical to SEQ ID NO: 14574. In certain embodiments, one transposon end comprises at least 14, at least 16, at least 18, at least 20 or at least 25 contiguous nucleotides from SEQ ID NO: 14573 and one transposon end comprises at least 14, at least 16, at least 18, at least 20 or at least 25 contiguous nucleotides from SEQ ID NO: 14574. In certain embodiments, one transposon end comprises at least 14, at least 16, at least 18, at least 20 from SEQ ID NO: 14591, and the other end comprises at least 14, at least 16, at least 18, at least 20 from SEQ ID NO: 14593. In certain embodiments, each transposon end comprises SEQ ID NO: 14575 in inverted orientations.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence selected from of SEQ ID NO: 14573, SEQ ID NO: 14579, SEQ ID NO: 14581, SEQ ID NO: 14582, SEQ ID NO: 14583, and SEQ ID NO: 14588, and a sequence selected from SEQ ID NO: 14587, SEQ ID NO: 14588, SEQ ID NO: 14589 and SEQ ID NO: 14586 and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14517 or SEQ ID NO: 14518.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises ITRs of CCCTTTGCCTGCCA (SEQ ID NO: 14622) (left ITR) and TGGCAGTGAAAGGG (SEQ ID NO: 14623) (right ITR) adjacent to the target sequences.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Helicoverpa armigera. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14525)

1
MASRQRLNHD EIATILENDD DYSPLDSESE KEDCVVEDDV WSDNEDAIVD FVEDTSAQED

61
PDNNIASRES PNLEVTSLTS HRIITLPQRS IRGKNNHVWS TTKGRTTGRT SAINIIRTNR

121
GPTRMCRNIV DPLLCFQLFI TDEIIHEIVK WTNVEIIVKR QNLKDISASY RDINTMEIWA

181
LVGILTLTAV MKDNHLSTDE LFDATFSGTR YVSVMSRERF EFLIRCIRMD DKTLRPTLRS

241
DDAFLPVRKI WEIFINQCRQ NHVPGSNLTV DEQLLGFRGR CPFRMYIPNK PDKYGIKFPM

301
MCAAATKYMI DAIPYLGKST KTNGLPLGEF YVKDLTKTVH GTNRNITCDN WFTSIPLAKN

361
MLQAPYNLTI VGTIRSNKRE MPEEIKNSRS RPVGSSMFCF DGPLTLVSYK PKPSKMVFLL

421
SSCDENAVIN ESNGKPDMIL FYNQTKGGVD SFDQMCKSMS ANRKTNRWPM AVFYGMLNMA

481
FVNSYIIYCH NKINKQEKPI SRKEFMKKLS IQLTTPWMQE RLQAPTLKRT LRDNITNVLK

541
NVVPASSENI SNEPEPKKRR YCGVCSYKKR RMTKAQCCKC KKAICGEHNI DVCQDCI.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Helicoverpa armigera. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14570)

1
ttaaccctag aagcccaatc tacgtaaatt tgacgtatac cgcggcgaaa tatctctgtc

61
tctttcatgt ttaccgtcgg atcgccgcta acttctgaac caactcagta gccattggga

121
cctcgcagga cacagttgcg tcatctcggt aagtgccgcc attttgttgt actctctatt

181
acaacacacg tcacgtcacg tcgttgcacg tcattttgac gtataattgg gctttgtgta

241
acttttgaat ttgtttcaaa ttttttatgt ttgtgattta tttgagttaa tcgtattgtt

301
tcgttacatt tttcatataa taataatatt ttcaggttga gtacaaa.

14570). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14528)

1
agactgtttt tttctaagag acttctaaaa tattattacg agttgattta attttatgaa

61
aacatttaaa actagttgat tttttttata attacataat tttaagaaaa agtgttagag

121
gcttgatttt tttgttgatt ttttctaaga tttgattaaa gtgccataat agtattaata

181
aagagtattt tttaacttaa aatgtatttt atttattaat taaaacttca attatgataa

241
ctcatgcaaa aatatagttc attaacagaa aaaaatagga aaactttgaa gttttgtttt

301
tacacgtcat ttttacgtat gattgggctt tatagctagt taaatatgat tgggcttcta

361
gggttaa .

in certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Pectinophora gossypiella. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14530)

1
MDLRKQDEKI RQWLEQDIEE DSKGESDNSS SETEDIVEME VHKNTSSESE VSSESDYEPV

61
CPSKRQRTQI IESEESDNSE SIRPSRRQTS RVIDSDETDE DVMSSTPQNI PRNPNVIQPS

121
SRFLYGKNKH KWSSAAKPSS VRTSRRNIIH FIPGPKERAR EVSEPIDIFS LFISEDMLQQ

181
VVTFTNAEML IRKNKYKTET FTVSPTNLEE IRALLGLLFN AAAMKSNHLP TRMLFNTHRS

241
GTIFKACMSA ERLNFLIKCL RFDDKLTRNV RQRDDRFAPI RDLWQALISN FQKWYTPGSY

301
ITVDEQLVGF RGRCSFRMYI PNKPNKYGIK LVMAADVNSK YIVNAIPYLG KGTDPQNQPL

361
ATFFIKEITS TLHGTNRNIT MDNWFTSVPL ANELLMAPYN LTLVGTLRSN KREIPEKLKN

421
SKSRAIGTSM FCYDGDKTLV SYKAKSNKVV FILSTIHDQP DINQETGKPE MIHFYNSTKG

481
AVDTVDQMCS SISTNRKTQR WPLCVFYNML NLSIINAYVV YVYNNVRNNK KPMSRRDFVI

541
KLGDQLMEPW LRQRLQTVTL RRDIKVMIQD ILGESSDLEA PVPSVSNVRK IYYLCPSKAR

601
RMTKHRCIKC KQAICGPHNI DICSRCIE.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Pectinophora gossypiella. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14532)

1
ttaaccctag ataactaaac attcgtccgc tcgacgacgc gctatgccgc gaaattgaag

61
tttacctatt attccgcgtc ccccgccccc gccgcttttt ctagcttcct gatttgcaaa

121
atagtgcatc gcgtgacacg ctcgaggtca cacgacaatt aggtcgaaag ttacaggaat

181
ttcgtcgtcc gctcgacgaa agtttagtaa ttacgtaagt ttggcaaagg taagtgaatg

241
aagtattttt ttataattat tttttaattc tttatagtga taacgtaagg tttatttaaa

301
tttattactt ttatagttat ttagccaatt gttataaatt ccttgttatt gctgaaaaat

361
ttgcctgttt tagtcaaaat ttattaactt ttcgatcgtt ttttag.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14571)

1
tttcactaag taattttgtt cctatttagt agataagtaa cacataatta ttgtgatatt

61
caaaacttaa gaggtttaat aaataataat aaaaaaaaaa tggtttttat ttcgtagtct

121
gctcgacgaa tgtttagtta ttacgtaacc gtgaatatag tttagtagtc tagggttaa.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Ctenoplusia agnata. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14534)

1
MASRQHLYQD EIAAILENED DYSPHDTDSE MEDCVTQDDV RSDVEDEMVD NIGNGTSPAS

61
RHEDPETPDP SSEASNLEVT LSSHRIIILP QRSIREKNNH IWSTTKGQSS GRTAAINIVR

121
TNRGPTRMCR NIVDPLLCFQ LFIKEEIVEE IVKWTNVEMV QKRVNLKDIS ASYRDTNEME

181
IWAIISMLTL SAVMKDNHLS TDELFNVSYG TRYVSVMSRE RFEFLLRLLR MGDKLLRPNL

241
RQEDAFTPVR KIWEIFINQC RLNYVPGTNL TVDEQLLGFR GRCPFRMYIP NKPDKYGIKF

301
PMVCDAATKY MVDAIPYLGK STKTQGLPLG EFYVKELTQT VHGTNRNVTC DNWFTSVPLA

361
KSLLNSPYNL TLVGTIRSNK REIPEEVKNS RSRQVGSSMF CFDGPLTLVS YKPKPSKMVF

421
LLSSCNEDAV VNQSNGKPDM ILFYNQTKGG VDSFDQMCSS MSTNRKTNRW PMAVFYGMLN

481
MAFVNSYIIY CHNMLAKKEK PLSRKDFMKK LSTDLTTPSM QKRLEAPTLK RSLRDNITNV

541
LKIVPQAAID TSFDEPEPKK RRYCGFCSYK KKRMTKTQCF KCKKPVCGEH NIDVCQDCI.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Ctenoplusia agnata. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14535)

1
ttaaccctag aagcccaatc tacgtcattc tgacgtgtat gtcgccgaaa atactctgtc

61
tctttctcct gcacgatcgg attgccgcga acgctcgatt caacccagtt ggcgccgaga

121
tctattggag gactgcggcg ttgattcggt aagtcccgcc attttgtcat agtaacagta

181
ttgcacgtca gcttgacgta tatttgggct ttgtgttatt tttgtaaatt ttcaacgtta

241
gtttattatt gcatcttttt gttacattac tggtttattt gcatgtatta ctcaaatatt

301
atttttattt tagcgtagaa aataca.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14536)

1 agactgtttt ttttgtattt gcattatata

ttatattcta aagttgattt aattctaaga

61 aaaacattaa aataagtttc tttttgtaaa

atttaattaa ttataagaaa aagtttaagt

121 tgatctcatt ttttataaaa atttgcaatg

tttccaaagt tattattgta aaagaataaa

181 taaaagtaaa ctgagtttta attgatgttt

tattatatca ttatactata tattacttaa

241 ataaaacaat aactgaatgt atttctaaaa

ggaatcacta gaaaatatag tgatcaaaaa

301 tttacacgtc atttttgcgt atgattgggc

tttataggtt ctaaaaatat gattgggcct

361 ctagggttaa.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of CCCTAGAAGCCCAATC (SEQ ID NO: 14564).

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Agrotis ipsilon. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14537)

1 MESRQRLNQD EIATILENDD DYSPLDSDSE

AEDRVVEDDV WSDNEDAMID YVEDTSRQED

61 PDNNIASQES ANLEVTSLTS HRIISLPQRS

ICGKNNHVWS TTKGRTTGRT SAINIIRTNR

121 GPTRMCRNIV DPLLCFQLFI TDEIIHEIVK

WTNVEMIVKR QNLIDISASY RDTNTMEMWA

181 LVGILTLTAV MKDNHLSTDE LFDATFSGTR

YVSVMSRERF EFLIRCMRMD DKTLRPTLRS

241 DDAFIPVRKL WEIFINQCRL NYVPGGNLTV

DEQLLGFRGR CPFRMYIPNK PDKYGIRFPM

301 MCDAATKYMI DAIPYLGKST KTNGLPLGEF

YVKELTKTVH GTNRNVTCDN WFTSIPLAKN

361 MLQAPYNLTI VGTIRSNKRE IPEEIKNSRS

RPVGSSMFCF DGPLTLVSYK PKPSRMVFLL

421 SSCDENAVIN ESNGKPDMIL FYNQTKGGVD

SFDQMCKSMS ANRKTNRWPM AVFYGMLNMA

481 FVNSYIIYCH NKINKQKKPI NRKEFMKNLS

TDLTTPWMQE RLKAPTLKRT LRDNITNVLK

541 NVVPPSPANN SEEPGPKKRS YCGFCSYKKR

RMTKTQFYKC KKAICGEHNI DVCQDCV.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Agrotis ipsilon. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14538)

1 ttaaccctag aagcccaatc tacgtaaatt

tgacgtatac cgcggcgaaa tatatctgtc

61 tctttcacgt ttaccgtcgg attcccgcta

acttcggaac caactcagta gccattgaga

121 actcccagga cacagttgcg tcatctcggt

aagtgccgcc attttgttgt aatagacagg

181 ttgcacgtca ttttgacgta taattgggct

ttgtgtaact tttgaaatta tttataattt

241 ttattgatgt gatttatttg agttaatcgt

attgtttcgt tacatttttc atatgatatt

301 aatattttca gattgaatat aaa.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14539)

1 agactgtttt ttttaaaagg cttataaagt

attactattg cgtgatttaa ttttataaaa

61 atatttaaaa ccagttgatt tttttaataa

ttacctaatt ttaagaaaaa atgttagaag

121 cttgatattt ttgttgattt ttttctaaga

tttgattaaa aggccataat tgtattaata

181 aagagtattt ttaacttcaa atttatttta

tttattaatt aaaacttcaa ttatgataat

241 acatgcaaaa atatagttca tcaacagaaa

aatataggaa aactctaata gttttatttt

301 tacacgtcat ttttacgtat gattgggctt

tatagctagt caaatatgat tgggcttcta

361 gggttaa.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Megachile rotundata. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14540)

1 MNGKDSLGEF YLDDLSDCLD CRSASSTDDE

SDSSNIAIRK RCPIPLIYSD SEDEDMNNNV

61 EDNNHFVKES NRYHYQIVEK YKITSKTKKW

KDVTVTEMKK FLGLIILMGQ VKKDVLYDYW

121 STDPSIETPF FSKVMSRNRF LQIMQSWHFY

NNNDISPNSH RLVKIQPVID YFKEKFNNVY

181 KSDQQLSLDE CLIPWRGRLS IKTYNPAKIT

KYGILVRVLS EARTGYVSNF CVYAADGKKI

241 EETVLSVIGP YKNMWHHVYQ DNYYNSVNIA

KIFLKNKLRV CGTIRKNRSL PQILQTVKLS

301 RGQHQFLRNG HTLLEVWNNG KRNVNMISTI

HSAQMAESRN RSRTSDCPIQ KPISIIDYNK

361 YMKGVDRADQ YLSYYSIFRK TKKWTKRVVM

FFINCALFNS FKVYTTLNGQ KITYKNFLHK

421 AALSLIEDCG TEEQGTDLPN SEPTTTRTTS

RVDHPGRLEN FGKHKLVNIV TSGQCKKPLR

481 QCRVCASKKK LSRTGFACKY CNVPLHKGDC

FERYHSLKKY.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Megachile rotundata. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14541)

1 ttaaataatg cccactctag atgaacttaa

cactttaccg accggccgtc gattattcga

61 cgtttgctcc ccagcgctta ccgaccggcc

atcgattatt cgacgtttgc ttcccagcgc

121 ttaccgaccg gtcatcgact tttgatcttt

ccgttagatt tggttaggtc agattgacaa

181 gtagcaagca tttcgcattc tttattcaaa

taatcggtgc ttttttctaa gctttagccc

241 ttagaa.

In certain embodiments, the the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14542)

1 acaacttctt ttttcaacaa atattgttat

atggattatt tatttattta tttatttatg

61 gtatatttta tgtttattta tttatggtta

ttatggtata ttttatgtaa ataataaact

121 gaaaacgatt gtaatagatg aaataaatat

tgttttaaca ctaatataat taaagtaaaa

181 gattttaata aatttcgtta ccctacaata

acacgaagcg tacaatttta ccagagttta

241 ttaa.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Bombus impatiens. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14543)

1 MNEKNGIGEF YLDDLSDCPD SYSRSNSGDE

SDGSDTIIRK RGSVLPPRYS DSEDDEINNV

61 EDNANNVENN DDIWSTNDEA IILEPFEGSP

GLKIMPSSAE SVTDNVNLFF GDDFFEHLVR

121 ESNRYHYQVM EKYKIPSKAK KWTDITVPEM

KKFLGLIVLM GQIKKDVLYD YWSTDPSIET

181 PFFSQVMSRN RFVQIMQSWH FCNNDNIPHD

SHRLAKIQPV IDYFRRKFND VYKPCQQLSL

241 DESIIPWRGR LSIKTYNPAK ITKYGILVRV

LSEAVTGYVC NFDVYAADGK KLEDTAVIEP

301 YKNIWHQIYQ DNYYNSVKMA RILLKNKVRV

CGTIRKNRGL PRSLKTIQLS RGQYEFRRNH

361 QILLEVWNNG RRNVNMISTI HSAQLMESRS

KSKRSDVPIQ KPNSIIDYNK YMKGVDRADQ

421 YLAYYSIFRK TKKWTKRVVM FFINCALFNS

FRVYTILNGK NITYKNFLHK VAVSWIEDGE

481 TNCTEQDDNL PNSEPTRRAP RLDHPGRLSN

YGKHKLINIV TSGRSLKPQR QCRVCAVQKK

541 RSRTCFVCKF CNVPLHKGDC FERYHTLKKY.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Bombus impatiens. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14544)

1 ttaatttttt aacattttac cgaccgatag

ccgattaatc gggtttttgc cgctgacgct

61 taccgaccga taacctatta atcggctttt

tgtcgtcgaa gcttaccaac ctatagccta

121 cctatagtta atcggttgcc atggcgataa

acaatctttc tcattatatg agcagtaatt

181 tgttatttag tactaaggta ccttgctcag

ttgcgtcagt tgcgttgctt tgtaagctcc

241 cacagtttta taccaattcg aaaaacttac

cgttcgcg.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14545)

1 actatttcac atttgaacta aaaaccgttg

taatagataa aataaatata atttagtatt

61 aatattatgg aaacaaaaga ttttattcaa

tttaattatc ctatagtaac aaaaagcggc

121 caattttatc tgagcatacg aaaagcacag

atactcccgc ccgacagtct aaaccgaaac

181 agagccggcg ccagggagaa tctgcgcctg

agcagccggt cggacgtgcg tttgctgttg

241 aaccgctagt ggtcagtaaa ccagaaccag

tcagtaagcc agtaactgat cagttaacta

301 gattgtatag ttcaaattga acttaatcta

gtttttaagc gtttgaatgt tgtctaactt

361 cgttatatat tatattcttt ttaa.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Mamestra brassicae. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14546)

1 MFSFVPNKEQ TRTVLIFCFH LKTTAAESHR

PLVEAFGEQV PTVKTCERWF QRFKSGDFDV

61 DDKEHGKPPK RYEDAELQAL LDEDDAQTQK

QLAEQLEVSQ QAVSNRLREG GKIQKVGRWV

121 PHELNERQRE RRKNTCEILL SRYKRKSFLH

RIVTGEEKWI FFVNPKRKKS YVDPGQPATS

181 TARPNRFGKK TRLCVWWDQS GVIYYELLKP

GETVNTARYQ QQLINLNRAL QRKRPEYQKR

241 QHRVIFLHDN APSHTARAVR DTLETLNWEV

LPHAAYSPDL APSDYHLFAS MGHALAEQRF

301 DSYESVEEWL DEWFAAKDDE FYWRGIHKLP

ERWDNCVASD GKYFE.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Mamestra brassicae. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14547)

1 ttattgggtt gcccaaaaag taattgcgga

tttttcatat acctgtcttt taaacgtaca

61 tagggatcga actcagtaaa actttgacct

tgtgaaataa caaacttgac tgtccaacca

121 ccatagtttg gcgcgaattg agcgtcataa

ttgttttgac tttttgcagt caac.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14548)

1 atgatttttt ctttttaaac caattttaat

tagttaattg atataaaaat ccgcaattac

61 tttttgggca acccaataa.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Mayetiola destructor. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14549)

1 MENFENWRKR RHLREVLLGH FFAKKTAAES

HRLLVEVYGE HALAKTQCFE WFQRFKSGDF

61 DTEDKERPGQ PKKFEDEELE ALLDEDCCQT

QEELAKSLGV TQQAISKRLK AAGYIQKQGN

121 WVPHELKPRD VERRFCMSEM LLQRHKKKSF

LSRIITGDEK WIHYDNSKRK KSYVKRGGRA

181 KSTPKSNLHG AKVMLCIWWD QRGVLYYELL

EPGQTITGDL YRTQLIRLKQ ALAEKRPEYA

241 KRHGAVIFHH DNARPHVALP VKNYLENSGW

EVLPHPPYSP DLAPSDYHLF RSMQNDLAGK

301 RFTSEQGIRK WLDSFLAAKP AKFFEKGIHE

LSERWEKVIA SDGQYFE.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Mayetiola destructor. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14550)

1 taagacttcc aaaatttcca cccgaacttt

accttccccg cgcattatgt ctctcttttc

61 accctctgat ccctggtatt gttgtcgagc

acgatttata ttgggtgtac aacttaaaaa

121 ccggaattgg acgctagatg tccacactaa

cgaatagtgt aaaagcacaa atttcatata

181 tacgtcattt tgaaggtaca tttgacagct

atcaaaatca gtcaataaaa ctattctatc

241 tgtgtgcatc atattttttt attaact.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14551)

1
tgcattcatt cattttgtta tcgaaataaa gcattaattt tcactaaaaa attccggttt

61
ttaagttgta cacccaatat catccttagt gacaattttc aaatggcttt cccattgagc

121
tgaaaccgtg gctctagtaa gaaaaacgcc caacccgtca tcatatgcct tttttttctc

181
aacatccg.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Apis mellifera. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14552)

1
MENQKEHYRH ILLFYFRKGK NASQAHKKLC AVYGDEALKE RQCQNWFDKF RSGDFSLKDE

61
KRSGRPVEVD DDLIKAIIDS DRHSTTREIA EKLHVSHTCI ENHLKQLGYV QKLDTWVPHE

121
LKEKHLTQRI NSCDLLKKRN ENDPFLKRLI TGDEKWVVYN NIKRKRSWSR PREPAQTTSK

181
AGIHRKKVLL SVWWDYKGIV YFELLPPNRT INSVVYIEQL TKLNNAVEEK RPELTNRKGV

241
VFHHDNARPH TSLVTRQKLL ELGWDVLPHP PYSPDLAPSD YFLFRSLQNS LNGKNFNNDD

301
DIKSYLIQFF ANKNQKFYER GIMMLPERWQ KVIDQNGQHI TE.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Apis mellifera. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14553)

1
ttgggttggc aactaagtaa ttgcggattt cactcataga tggcttcagt tgaattttta

61
ggtttgctgg cgtagtccaa atgtaaaaca cattttgtta tttgatagtt ggcaattcag

121
ctgtcaatca gtaaaaaaag ttttttgatc ggttgcgtag ttttcgtttg gcgttcgttg

181
aaaa.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14554)

1
agttatttag ttccatgaaa aaattgtctt tgattttcta aaaaaaatcc gcaattactt

61
agttgccaat ccaa.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Messor bouvieri. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14555)

1
MSSFVPENVH LRHALLFLFH QKKRAAESHR LLVETYGEHA PTIRTCETWF RQFKCGDFNV

61
QDKERPGRPK TFEDAELQEL LDEDSTQTQK QLAEKLNVSR VAICERLQAM GKIQKMGRWV

121
PHELNDRQME NRKIVSEMLL QRYERKSFLH RIVTGDEKWI YFENPKRKKS WLSPGEAGPS

181
TARPNRFGRK TMLCVWWDQI GVVYYELLKP GETVNTDRYR QQMINLNCAL IEKRPQYAQR

241
HDKVILQHDN APSHTAKPVK EMLKSLGWEV LSHPPYSPDL APSDYHLFAS MGHALAEQHF

301
ADFEEVKKWL DEWFSSKEKL FFWNGIHKLS ERWTKCIESN GQYFE.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Messor bouvieri. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14556)

1
agtcagaaat gacacctcga tcgacgacta atcgacgtct aatcgacgtc gattttatgt

61
caacatgtta ccaggtgtgt cggtaattcc tttccggttt ttccggcaga tgtcactagc

121
cataagtatg aaatgttatg atttgataca tatgtcattt tattctactg acattaacct

181
taaaactaca caagttacgt tccgccaaaa taacagcgtt atagatttat aattttttga

241
aa.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14557)

1
ataaatttga actatccatt ctaagtaacg tgttttcttt aacgaaaaaa ccggaaaaga

61
attaccgaca ctcctggtat gtcaacatgt tattttcgac attgaatcgc gtcgattcga

121
agtcgatcga ggtgtcattt ctgact.

In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Trichoplusia ni. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

(SEQ ID NO: 14558)

1
MGSSLDDEHI LSALLQSDDE LVGEDSDSEV SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61
SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121
PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTSATFRD TNEDEIYAFF

181
GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV

241
FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RVYIPNKPSK YGIKILMMCD

301
SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ

361
EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC

421
DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481
SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPKEV

541
PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Trichoplusia ni. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14559)

1
ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc

61
tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga

121
gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc

181
gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg

241
ataattatat tgttatttca tgttctactt acgtgataac ttattatata tatattttct

301
tgttatagat atc.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14560)

1
tttgttactt tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat

61
aaataaattg tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat

121
atctattcaa attaataaat aaacctcgat atacagaccg ataaaacaca tgcgtcaatt

181
ttacgcatga ttatctttaa cgtacgtcac aatatgatta tctttctagg gttaa.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14561)

1
ccctagaaag atagtctgcg taaaattgac gcatgcattc ttgaaatatt gctctctctt

61
tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg cttggagctc

121
ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata acgaccgcgt

181
gagtcaaaat gacgcatgat tatcttttac gtgactttta agatttaact catacgataa

241
ttatattgtt atttcatgtt ctacttacgt gataacttat tatatatata ttttcttgtt

301
atagatatc.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14562)

1
tttgttactt tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat

61
aaataaattg tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat

121
atctattcaa attaataaat aaacctcgat atacagaccg ataaaacaca tgcgtcaatt

181
ttacgcatga ttatctttaa cgtacgtcac aatatgatta tctttctagg g.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14609)

1
tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg cttggagctc

61
ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata acgaccgcgt

121
gagtcaaaat gacgcatgat tatcttttac gtgactttta agatttaact catacgataa

181
ttatattgtt atttcatgtt ctacttacgt gataacttat tatatatata ttttcttgtt

241
atagatatc.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

(SEQ ID NO: 14610)

1
tttgttactt tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat

61
aaataaattg tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat

121
atctattcaa attaataaat aaacctcgat atacagaccg ataaaacaca tgcgtcaatt

181
ttacgcatga ttatctttaa cgtacgtcac aatatgatta tctttctagg g.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14561 and SEQ ID NO: 14562, and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14558. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14609 and SEQ ID NO: 14610, and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14558.

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Aphis gossypii. In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of CCTTCCAGCGGGCGCGC (SEQ ID NO: 14565).

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Chilo suppressalis. In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of CCCAGATTAGCCT (SEQ ID NO: 14566).

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Heliothis virescens. In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of CCCTTAATTACTCGCG (SEQ ID NO: 14567).

In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Anopheles stephensi. In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of CCCTAGAAAGATA (SEQ ID NO: 14569).

Gene Editing

In various embodiments, nucleases that may be used as cutting enzymes include, but are not limited to, Cas9, transcription activator-like effector nucleases (TALENs) and zinc finger nucleases. In certain embodiments, the Cas9 is a catalytically inactive or “inactivated” Cas9 (dCas9). In certain embodiments, the Cas9 is a catalytically inactive or “inactivated” nuclease domain of Cas9. In certain embodiments, the dCas9 is encoded by a shorter sequence that is derived from a full length, catalytically inactivated, Cas9, referred to herein as a “small” dCas9 or dSaCas9.

In certain embodiments, the inactivated, small, Cas9 (dSaCas9) operatively-linked to an active nuclease. In certain embodiments, the disclosure provides a fusion protein comprising, consisting essentially of or consisting of a DNA binding domain and molecule nuclease, wherein the nuclease comprises a small, inactivated Cas9 (dSaCas9). In certain embodiments, the dSaCas9 of the disclosure comprises the mutations D10A and N580A (underlined and bolded) which inactivate the catalytic site. In certain embodiments, the dSaCas9 (isolated or derived from Staphylococcus aureus) of the disclosure comprises the amino acid sequence of:

In certain embodiments of the gene editing systems of the disclosure, the dCas9 of the disclosure comprises a dCas9 isolated or derived from Streptococcus pyogenes. In certain embodiments, the dCas9 comprises a dCas9 with substitutions at positions 10 and 840 of the amino acid sequence of the dCas9 which inactivate the catalytic site. In certain embodiments, these substitutions are D10A and H840A. In certain embodiments, the amino acid sequence of the dCas9 (isolated or derived from Streptococcus pyogenes) comprises the sequence of:

(SEQ ID NO: 14498)

1 XDKKYSIGLA IGTNSVGWAV ITDEYKVPSK KFKVLGNTDR HSIKKNLIGA LLFDSGETAE

61 ATRLKRTARR RYTRRKNRIC YLQEIFSNEM AKVDDSFFHR LEESFLVEED KKHERHPIFG

121 NIVDEVAYHE KYPTIYHLRK KLVDSTDKAD LRLIYLALAH MIKFRGHFLI EGDLNPDNSD

181 VDKLFIQLVQ TYNQLFEENP INASGVDAKA ILSARLSKSR RLENLIAQLP GEKKNGLFGN

241 LIALSLGLTP NFKSNFDLAE DAKLQLSKDT YDDDLDNLLA QIGDQYADLF LAAKNLSDAI

301 LLSDILRVNT EITKAPLSAS MIKRYDEHHQ DLTLLKALVR QQLPEKYKEI FFDQSKNGYA

361 GYIDGGASQE EFYKFIKPIL EKMDGTEELL VKLNREDLLR KQRTFDNGSI PHQIHLGELH

421 AILRRQEDFY PFLKDNREKI EKILTFRIPY YVGPLARGNS RFAWMTRKSE ETITPWNFEE

481 VVDKGASAQS FIERMTNFDK NLPNEKVLPK HSLLYEYFTV YNELTKVKYV TEGMRKPAFL

541 SGEQKKAIVD LLFKTNRKVT VKQLKEDYFK KIECFDSVEI SGVEDRFNAS LGTYHDLLKI

601 IKDKDFLDNE ENEDILEDIV LTLTLFEDRE MIEERLKTYA HLFDDKVMKQ LKRRRYTGWG

661 RLSRKLINGI RDKQSGKTIL DFLKSDGFAN RNFMQLIHDD SLTFKEDIQK AQVSGQGDSL

721 HEHIANLAGS PAIKKGILQT VKVVDELVKV MGRHKPENIV IEMARENQTT QKGQKNSRER

781 MKRIEEGIKE LGSQILKEHP VENTQLQNEK LYLYYLQNGR DMYVDQELDI NRLSDYDVDA

841 IVPQSFLKDD SIDNKVLTRS DKNRGKSDNV PSEEVVKKMK NYWRQLLNAK LITQRKFDNL

901 TKAERGGLSE LDKAGFIKRQ LVETRQITKH VAQILDSRMN TKYDENDKLI REVKVITLKS

961 KLVSDFRKDF QFYKVREINN YHHAHDAYLN AVVGTALIKK YPKLESEFVY GDYKVYDVRK

1021 MIAKSEQEIG KATAKYFFYS NIMNFFKTEI TLANGEIRKR PLIETNGETG EIVWDKGRDF

1081 ATVRKVLSMP QVNIVKKTEV QTGGFSKESI LPKRNSDKLI ARKKDWDPKK YGGFDSPTVA

1141 YSVLVVAKVE KGKSKKLKSV KELLGITIME RSSFEKNPID FLEAKGYKEV KKDLIIKLPK

1201 YSLFELENGR KRMLASAGEL QKGNELALPS KYVNFLYLAS HYEKLKGSPE DNEQKQLFVE

1261 QHKHYLDEII EQISEFSKRV ILADANLDKV LSAYNKHRDK PIREQAENII HLFTLTNLGA

1321 PAAFKYFDTT IDRKRYTSTK EVLDATLIHQ SITGLYETRI DLSQLGGD.

In certain embodiments of the gene editing systems of the disclosure, the nuclease domain may comprise, consist essentially of or consist of a dCas9 or a dSaCas9 and a type IIS endonuclease. In certain embodiments of the disclosure, the nuclease domain may comprise, consist essentially of or consist of a dSaCas9 and a type IIS endonuclease, including, but not limited to, AciI, Mn1I, AlwI, BbvI, BccI, BceAI, BsmAI, BsmFI, BspCNI, BsrI, BtsCI, HgaI, HphI, HpyAV, MbolI, My1I, PleI, SfaNI, AcuI, BciVI, BfuAI, BmgBI, BmrI, BpmI, BpuEI, BsaI, BseRI, BsgI, BsmI, BspMI, BsrBI, BsrBI, BsrDI, BtgZI, BtsI, EarI, EciI, MmeI, NmeAIII, BbvCI, Bpu10I, BspQI, SapI, BaeI, BsaXI, CspCI, BfiI, MboII, Acc36I, FokI or Clo051. In certain embodiments of the disclosure, the nuclease domain may comprise, consist essentially of or consist of a dSaCas9 and Clo051. An exemplary Clo051 nuclease domain may comprise, consist essentially of or consist of, the amino acid sequence of:

(SEQ ID NO: 14503)

EGIKSNISLLKDELRGQISHISHEYLSLIDLAFDSKQNRLFEMKVLELLV

NEYGFKGRHLGGSRKPDGIVYSTTLEDNFGIIVDTKAYSEGYSLPISQAD

EMERYVRENSNRDEEVNPNKWWENFSEEVKKYYFVFISGSFKGKFEEQLR

RLSMTTGVNGSAVNVVNLLLGAEKIRSGEMTIEELERAMFNNSEFILKY.

An exemplary dCas9-Clo051 nuclease domain may comprise, consist essentially of or consist of, the amino acid sequence of (Clo051 sequence underlined, linker bold italics, dCas9 (Staphylococcus pyogenes) sequence in italics):

(SEQ ID NO: 14654)

MAPKKKRKVEGIKSNISLLKDELRGQISHISHEYLSLIDLAFDSKQNRLF

EMKVLELLVNEYGFKGRHLGGSRKPDGIVYSTTLEDNFGIIVDTKAYSEG

YSLPISQADEMERYVRENSNRDEEVNPNKWWENFSEEVKKYYFVFISGSF

KGKFEEQLRRLSMTTGVNGSAVNVVNLLLGAEKIRSGEMTIEELERAMFN

NSEFILKY
custom-character

DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNT

DRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSN

EIVIAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIY

HLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFI

QLVQTYNQLFEENPINASGVDAICAILSARLSKSRRLENLIAQLPGEKKN

GLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQ

YADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLL

KALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEICMD

GTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLK

DNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDK

GASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGM

RKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE

DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREAKEE

RLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLK

SDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIK

KGILQTVKVVDELVKVMGRHKPENIVIEIVIARENQTTQKGQKNSRERMI

CRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDIN

RLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKN

YWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV

AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNY

HHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRICAKAKSEQEIG

KATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF

ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKK

YGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPID

FLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPS

KYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEHEQISEFSKRVI

LADANLDKVLSAYNKHRDKPIREQAENIHILFTLTNLGAPAAFKYFDTTI

DRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDGSPKKKRKVSS.

Gene editing compositions of the disclosure may comprise a nuclease protein or a nuclease domain thereof. In certain embodiments, the gene editing composition comprises a sequence encoding a nuclease protein or a sequence encoding a nuclease domain thereof. In certain embodiments, the sequence encoding a nuclease protein or the sequence encoding a nuclease domain thereof comprises a DNA sequence, an RNA sequence, or a combination thereof. In certain embodiments, the nuclease or the nuclease domain thereof comprises one or more of a CRISPR/Cas protein, a Transcription Activator-Like Effector Nuclease (TALEN), a Zinc Finger Nuclease (ZFN), and an endonuclease. In certain embodiments, the nuclease or the nuclease domain thereof comprises one or more of a nuclease-inactivated Cas (dCas) protein, a Transcription Activator-Like Effector Nuclease (TALEN), a Zinc Finger Nuclease (ZFN), and an endonuclease. In certain embodiments, the nuclease or the nuclease domain thereof comprises a nuclease-inactivated Cas (dCas) protein and an endonuclease. In certain embodiments, the nuclease or the nuclease domain thereof comprises a nuclease-inactivated Cas9 (dCas9) protein and an endonuclease, wherein the endonuclease comprises a Clo051 nuclease or a nuclease domain thereof. In certain embodiments, the gene editing composition comprises a fusion protein. In certain embodiments, the fusion protein comprises a nuclease-inactivated Cas9 (dCas9) protein and a Clo051 nuclease or a Clo051 nuclease domain. In certain embodiments, the gene editing composition further comprises a guide sequence. In certain embodiments, the guide sequence comprises an RNA sequence.

In certain embodiments, the gene editing composition comprises a fusion protein. In certain embodiments, the fusion protein comprises a nuclease-inactivated Cas9 (dCas9) protein and a Clo051 nuclease or a Clo051 nuclease domain. In certain embodiments, the gene editing composition further comprises a guide sequence. In certain embodiments, the guide sequence comprises an RNA sequence. In certain embodiments, the fusion protein comprises or consists of the amino acid sequence:

(SEQ ID NO: 14654)

MAPKKKRKVEGIKSNISLLKDELRGQISHISHEYLSLIDLAFDSKQNRLF

EMKVLELLVNEYGFKGRHLGGSRKPDGIVYSTTLEDNEGIIVDTKAYSEG

YSLPISQADEMERYVRENSNRDEEVNPNKWWENFSEEVKKYYFVFISGSF

KGKFEEQLRRLSMTTGVNGSAVNVVNLLLGAEKIRSGEMTIEELERAMEN

NSEFILKYGGGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKEKVLGNT

DRHSIKKNLIGALLEDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSN

EMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHL

RKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQL

VQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF

GNLIALSLGLIPNEKSNEDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYAD

LFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKAL

VRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEE

LLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNRE

KIEKILTERIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASA

QSFIERMTNEDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPA

FLSGEQKKAIVDLLEKTNRKVIVKQLKEDYFKKIECEDSVEISGVEDRFN

ASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKT

YAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGF

ANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGIL

QTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGI

KELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDV

DAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLN

AKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSR

MNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAY

LNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFF

YSNIMNFEKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS

MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT

VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYK

EVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYL

ASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD

KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTS

TKEVLDATLIHQSITGLYETRIDLSQLGGDGSPKKKRKVSS.

In certain embodiments, the fusion protein is encoded by a nucleic acid comprising or consisting of the sequence:

(SEQ ID NO: 14655)

1 atggcaccaa agaagaaaag aaaagtggag ggcatcaagt caaacatcag cctgctgaaa

61 gacgaactgc ggggacagat tagtcacatc agtcacgagt acctgtcact gattgatctg

121 gccttcgaca gcaagcagaa tagactgttt gagatgaaag tgctggaact gctggtcaac

181 gagtatggct tcaagggcag acatctgggc gggtctagga aacctgacgg catcgtgtac

241 agtaccacac tggaagacaa cttcggaatc attgtcgata ccaaggctta ttccgagggc

301 tactctctgc caattagtca ggcagatgag atggaaaggt acgtgcgcga aaactcaaat

361 agggacgagg aagtcaaccc caataagtgg tgggagaatt tcagcgagga agtgaagaaa

421 tactacttcg tctttatctc aggcagcttc aaagggaagt ttgaggaaca gctgcggaga

481 ctgtccatga ctaccggggt gaacggatct gctgtcaacg tggtcaatct gctgctgggc

541 gcagaaaaga tcaggtccgg ggagatgaca attgaggaac tggaacgcgc catgttcaac

601 aattctgagt ttatcctgaa gtatggaggc gggggaagcg ataagaaata ctccatcgga

661 ctggccattg gcaccaattc cgtgggctgg gctgtcatca cagacgagta caaggtgcca

721 agcaagaagt tcaaggtcct ggggaacacc gatcgccaca gtatcaagaa aaatctgatt

781 ggagccctgc tgttcgactc aggcgagact gctgaagcaa cccgactgaa gcggactgct

841 aggcgccgat atacccggag aaaaaatcgg atctgctacc tgcaggaaat tttcagcaac

901 gagatggcca aggtggacga tagtttcttt caccgcctgg aggaatcatt cctggtggag

961 gaagataaga aacacgagcg gcatcccatc tttggcaaca ttgtggacga agtcgcttat

1021 cacgagaagt accctactat ctatcatctg aggaagaaac tggtggactc caccgataag

1081 gcagacctgc gcctgatcta tctggccctg gctcacatga tcaagttccg ggggcatttt

1141 ctgatcgagg gagatctgaa ccctgacaat tctgatgtgg acaagctgtt catccagctg

1201 gtccagacat acaatcagct gtttgaggaa aacccaatta atgcctcagg cgtggacgca

1261 aaggccatcc tgagcgccag actgtccaaa tctaggcgcc tggaaaacct gatcgctcag

1321 ctgccaggag agaagaaaaa cggcctgttt gggaatctga ttgcactgtc cctgggcctg

1381 acacccaact tcaagtctaa ttttgatctg gccgaggacg ctaagctgca gctgtccaaa

1441 gacacttatg acgatgacct ggataacctg ctggctcaga tcggcgatca gtacgcagac

1501 ctgttcctgg ccgctaagaa tctgagtgac gccatcctgc tgtcagatat tctgcgcgtg

1561 aacacagaga ttactaaggc cccactgagt gcttcaatga tcaaaagata tgacgagcac

1621 catcaggatc tgaccctgct gaaggctctg gtgaggcagc agctgcccga gaaatacaag

1681 gaaatcttct ttgatcagag caagaatgga tacgccggct atattgacgg cggggcttcc

1741 caggaggagt tctacaagtt catcaagccc attctggaaa agatggacgg caccgaggaa

1801 ctgctggtga agctgaatcg ggaggacctg ctgagaaaac agaggacatt tgataacgga

1861 agcatccctc accagattca tctgggcgaa ctgcacgcca tcctgcgacg gcaggaggac

1921 ttctacccat ttctgaagga taaccgcgag aaaatcgaaa agatcctgac cttcagaatc

1981 ccctactatg tggggcctct ggcacgggga aatagtagat ttgcctggat gacaagaaag

2041 tcagaggaaa ctatcacccc ctggaacttc gaggaagtgg tcgataaagg cgctagcgca

2101 cagtccttca ttgaaaggat gacaaatttt gacaagaacc tgccaaatga gaaggtgctg

2161 cccaaacaca gcctgctgta cgaatatttc acagtgtata acgagctgac taaagtgaag

2221 tacgtcaccg aagggatgcg caagcccgca ttcctgtccg gagagcagaa gaaagccatc

2281 gtggacctgc tgtttaagac aaatcggaaa gtgactgtca aacagctgaa ggaagactat

2341 ttcaagaaaa ttgagtgttt cgattcagtg gaaatcagcg gcgtcgagga caggtttaac

2401 gcctccctgg ggacctacca cgatctgctg aagatcatca aggataagga cttcctggac

2461 aacgaggaaa atgaggacat cctggaggac attgtgctga cactgactct gtttgaggat

2521 cgcgaaatga tcgaggaacg actgaagact tatgcccatc tgttcgatga caaagtgatg

2581 aagcagctga aaagaaggcg ctacaccgga tggggacgcc tgagccgaaa actgatcaat

2641 gggattagag acaagcagag cggaaaaact atcctggact ttctgaagtc cgatggcttc

2701 gccaacagga acttcatgca gctgattcac gatgactctc tgaccttcaa ggaggacatc

2761 cagaaagcac aggtgtctgg ccagggggac agtctgcacg agcatatcgc aaacctggcc

2821 ggcagccccg ccatcaagaa agggattctg cagaccgtga aggtggtgga cgaactggtc

2881 aaggtcatgg gacgacacaa acctgagaac atcgtgattg agatggcccg cgaaaatcag

2941 acaactcaga agggccagaa aaacagtcga gaacggatga agagaatcga ggaaggcatc

3001 aaggagctgg ggtcacagat cctgaaggag catcctgtgg aaaacactca gctgcagaat

3061 gagaaactgt atctgtacta tctgcagaat ggacgggata tgtacgtgga ccaggagctg

3121 gatattaaca gactgagtga ttatgacgtg gatgccatcg tccctcagag cttcctgaag

3181 gatgactcca ttgacaacaa ggtgctgacc aggtccgaca agaaccgcgg caaatcagat

3241 aatgtgccaa gcgaggaagt ggtcaagaaa atgaagaact actggaggca gctgctgaat

3301 gccaagctga tcacacagcg gaaatttgat aacctgacta aggcagaaag aggaggcctg

3361 tctgagctgg acaaggccgg cttcatcaag cggcagctgg tggagacaag acagatcact

3421 aagcacgtcg ctcagattct ggatagcaga atgaacacaa agtacgatga aaacgacaag

3481 ctgatcaggg aggtgaaagt cattactctg aaatccaagc tggtgtctga ctttagaaag

3541 gatttccagt tttataaagt cagggagatc aacaactacc accatgctca tgacgcatac

3601 ctgaacgcag tggtcgggac cgccctgatt aagaaatacc ccaagctgga gtccgagttc

3661 gtgtacggag actataaagt gtacgatgtc cggaagatga tcgccaaatc tgagcaggaa

3721 attggcaagg ccaccgctaa gtatttcttt tacagtaaca tcatgaattt ctttaagacc

3781 gaaatcacac tggcaaatgg ggagatcaga aaaaggcctc tgattgagac caacggggag

3841 acaggagaaa tcgtgtggga caagggaagg gattttgcta ccgtgcgcaa agtcctgtcc

3901 atgccccaag tgaatattgt caagaaaact gaagtgcaga ccgggggatt ctctaaggag

3961 agtattctgc ctaagcgaaa ctctgataaa ctgatcgccc ggaagaaaga ctgggacccc

4021 aagaagtatg gcgggttcga ctctccaaca gtggcttaca gtgtcctggt ggtcgcaaag

4081 gtggaaaagg ggaagtccaa gaaactgaag tctgtcaaag agctgctggg aatcactatt

4141 atggaacgca gctccttcga gaagaatcct atcgattttc tggaagccaa gggctataaa

4201 gaggtgaaga aagacctgat cattaagctg ccaaaatact cactgtttga gctggaaaac

4261 ggacgaaagc gaatgctggc aagcgccgga gaactgcaga agggcaatga gctggccctg

4321 ccctccaaat acgtgaactt cctgtatctg gctagccact acgagaaact gaaggggtcc

4381 cctgaggata acgaacagaa gcagctgttt gtggagcagc acaaacatta tctggacgag

4441 atcattgaac agatttcaga gttcagcaag agagtgatcc tggctgacgc aaatctggat

4501 aaagtcctga gcgcatacaa caagcaccga gacaaaccaa tccgggagca ggccgaaaat

4561 atcattcatc tgttcaccct gacaaacctg ggcgcccctg cagccttcaa gtattttgac

4621 accacaatcg atcggaagag atacacttct accaaagagg tgctggatgc taccctgatc

4681 caccagagta ttaccggcct gtatgagaca cgcatcgacc tgtcacagct gggaggcgat

4741 gggagcccca agaaaaagcg gaaggtgtct agttaa.

(SEQ ID NO: 14656)

1 MPKKKRKVEG IKSNISLLKD ELRGQISHIS HEYLSLIDLA FDSKQNRLFE MKVLELLVNE

61 YGFKGRHLGG SRKPDGIVYS TTLEDNFGII VDTKAYSEGY SLPISQADEM ERYVRENSNR

121 DEEVNPNKWW ENFSEEVKKY YFVFISGSFK GKFEEQLRRL SMTTGVNGSA VNVVNLLLGA

181 EKIRSGEMTI EELERAMFNN SEFILKYGGG GSDKKYSIGL AIGTNSVGWA VITDEYKVPS

241 KKFKVLGNTD RHSIKKNLIG ALLFDSGETA EATRLKRTAR RRYTRRKNRI CYLQEIFSNE

301 MAKVDDSFFH RLEESFLVEE DKKHERHPIF GNIVDEVAYH EKYPTIYHLR KKLVDSTDKA

361 DLRLIYLALA HMIKFRGHFL IEGDLNPDNS DVDKLFIQLV QTYNQLFEEN PINASGVDAK

421 AILSARLSKS RRLENLIAQL PGEKKNGLFG NLIALSLGLT PNFKSNFDLA EDAKLQLSKD

481 TYDDDLDNLL AQIGDQYADL FLAAKNLSDA ILLSDILRVN TEITKAPLSA SMIKRYDEHH

541 QDLTLLKALV RQQLPEKYKE IFFDQSKNGY AGYIDGGASQ EEFYKFIKPI LEKMDGTEEL

601 LVKLNREDLL RKQRTFDNGS IPHQIHLGEL HAILRRQEDF YPFLKDNREK IEKILTFRIP

661 YYVGPLARGN SRFAWMTRKS EETITPWNFE EVVDKGASAQ SFIERMTNFD KNLPNEKVLP

721 KHSLLYEYFT VYNELTKVKY VTEGMRKPAF LSGEQKKAIV DLLFKTNRKV TVKQLKEDYF

781 KKIECFDSVE ISGVEDRFNA SLGTYHDLLK IIKDKDFLDN EENEDILEDI VLTLTLFEDR

841 EMIEERLKTY AHLFDDKVMK QLKRRRYTGW GRLSRKLING IRDKQSGKTI LDFLKSDGFA

901 NRNFMQLIHD DSLTFKEDIQ KAQVSGQGDS LHEHIANLAG SPAIKKGILQ TVKVVDELVK

961 VMGRHKPENI VIEMARENQT TQKGQKNSRE RMKRIEEGIK ELGSQILKEH PVENTQLQNE

1021 KLYLYYLQNG RDMYVDQELD INRLSDYDVD AIVPQSFLKD DSIDNKVLTR SDKNRGKSDN

1081 VPSEEVVKKM KNYWRQLLNA KLITQRKFDN LTKAERGGLS ELDKAGFIKR QLVETRQITK

1141 HVAQILDSRM NTKYDENDKL IREVKVITLK SKLVSDFRKD FQFYKVREIN NYHHAHDAYL

1201 NAVVGTALIK KYPKLESEFV YGDYKVYDVR KMIAKSEQEI GKATAKYFFY SNIMNFFKTE

1261 ITLANGEIRK RPLIETNGET GEIVWDKGRD FATVRKVLSM PQVNIVKKTE VQTGGFSKES

1321 ILPKRNSDKL IARKKDWDPK KYGGFDSPTV AYSVLVVAKV EKGKSKKLKS VKELLGITIM

1381 ERSSFEKNPI DFLEAKGYKE VKKDLIIKLP KYSLFELENG RKRMLASAGE LQKGNELALP

1441 SKYVNFLYLA SHYEKLKGSP EDNEQKQLFV EQHKHYLDEI IEQISEFSKR VILADANLDK

1501 VLSAYNKHRD KPIREQAENI IHLFTLINLG APAAFKYFDT TIDRKRYTST KEVLDATLIH

1561 QSITGLYETR IDLSQLGGDG SPKKKRKV.

In certain embodiments, the fusion protein is encoded by a nucleic acid comprising or consisting of the sequence:

(SEQ ID NO: 14657)

1 atgcctaaga agaagcggaa ggtggaaggc atcaaaagca acatctccct cctgaaagac

61 gaactccggg ggcagattag ccacattagt cacgaatacc tctccctcat cgacctggct

121 ttcgatagca agcagaacag gctctttgag atgaaagtgc tggaactgct cgtcaatgag

181 tacgggttca agggtcgaca cctcggcgga tctaggaaac cagacggcat cgtgtatagt

241 accacactgg aagacaactt tgggatcatt gtggatacca aggcatactc tgagggttat

301 agtctgccca tttcacaggc cgacgagatg gaacggtacg tgcgcgagaa ctcaaataga

361 gatgaggaag tcaaccctaa caagtggtgg gagaacttct ctgaggaagt gaagaaatac

421 tacttcgtct ttatcagcgg gtccttcaag ggtaaatttg aggaacagct caggagactg

481 agcatgacta ccggcgtgaa tggcagcgcc gtcaacgtgg tcaatctgct cctgggcgct

541 gaaaagattc ggagcggaga gatgaccatc gaagagctgg agagggcaat gtttaataat

601 agcgagttta tcctgaaata cggtggcggt ggatccgata aaaagtattc tattggttta

661 gccatcggca ctaattccgt tggatgggct gtcataaccg atgaatacaa agtaccttca

721 aagaaattta aggtgttggg gaacacagac cgtcattcga ttaaaaagaa tcttatcggt

781 gccctcctat tcgatagtgg cgaaacggca gaggcgactc gcctgaaacg aaccgctcgg

841 agaaggtata cacgtcgcaa gaaccgaata tgttacttac aagaaatttt tagcaatgag

901 atggccaaag ttgacgattc tttctttcac cgtttggaag agtccttcct tgtcgaagag

961 gacaagaaac atgaacggca ccccatcttt ggaaacatag tagatgaggt ggcatatcat

1021 gaaaagtacc caacgattta tcacctcaga aaaaagctag ttgactcaac tgataaagcg

1081 gacctgaggt taatctactt ggctcttgcc catatgataa agttccgtgg gcactttctc

1141 attgagggtg atctaaatcc ggacaactcg gatgtcgaca aactgttcat ccagttagta

1201 caaacctata atcagttgtt tgaagagaac cctataaatg caagtggcgt ggatgcgaag

1261 gctattctta gcgcccgcct ctctaaatcc cgacggctag aaaacctgat cgcacaatta

1321 cccggagaga agaaaaatgg gttgttcggt aaccttatag cgctctcact aggcctgaca

1381 ccaaatttta agtcgaactt cgacttagct gaagatgcca aattgcagct tagtaaggac

1441 acgtacgatg acgatctcga caatctactg gcacaaattg gagatcagta tgcggactta

1501 tttttggctg ccaaaaacct tagcgatgca atcctcctat ctgacatact gagagttaat

1561 actgagatta ccaaggcgcc gttatccgct tcaatgatca aaaggtacga tgaacatcac

1621 caagacttga cacttctcaa ggccctagtc cgtcagcaac tgcctgagaa atataaggaa

1681 atattctttg atcagtcgaa aaacgggtac gcaggttata ttgacggcgg agcgagtcaa

1741 gaggaattct acaagtttat caaacccata ttagagaaga tggatgggac ggaagagttg

1801 cttgtaaaac tcaatcgcga agatctactg cgaaagcagc ggactttcga caacggtagc

1861 attccacatc aaatccactt aggcgaattg catgctatac ttagaaggca ggaggatttt

1921 tatccgttcc tcaaagacaa tcgtgaaaag attgagaaaa tcctaacctt tcgcatacct

1981 tactatgtgg gacccctggc ccgagggaac tctcggttcg catggatgac aagaaagtcc

2041 gaagaaacga ttactccatg gaattttgag gaagttgtcg ataaaggtgc gtcagctcaa

2101 tcgttcatcg agaggatgac caactttgac aagaatttac cgaacgaaaa agtattgcct

2161 aagcacagtt tactttacga gtatttcaca gtgtacaatg aactcacgaa agttaagtat

2221 gtcactgagg gcatgcgtaa acccgccttt ctaagcggag aacagaagaa agcaatagta

2281 gatctgttat tcaagaccaa ccgcaaagtg acagttaagc aattgaaaga ggactacttt

2341 aagaaaattg aatgcttcga ttctgtcgag atctccgggg tagaagatcg atttaatgcg

2401 tcacttggta cgtatcatga cctcctaaag ataattaaag ataaggactt cctggataac

2461 gaagagaatg aagatatctt agaagatata gtgttgactc ttaccctctt tgaagatcgg

2521 gaaatgattg aggaaagact aaaaacatac gctcacctgt tcgacgataa ggttatgaaa

2581 cagttaaaga ggcgtcgcta tacgggctgg ggacgattgt cgcggaaact tatcaacggg

2641 ataagagaca agcaaagtgg taaaactatt ctcgattttc taaagagcga cggcttcgcc

2701 aataggaact ttatgcagct gatccatgat gactctttaa ccttcaaaga ggatatacaa

2761 aaggcacagg tttccggaca aggggactca ttgcacgaac atattgcgaa tcttgctggt

2821 tcgccagcca tcaaaaaggg catactccag acagtcaaag tagtggatga gctagttaag

2881 gtcatgggac gtcacaaacc ggaaaacatt gtaatcgaga tggcacgcga aaatcaaacg

2941 actcagaagg ggcaaaaaaa cagtcgagag cggatgaaga gaatagaaga gggtattaaa

3001 gaactgggca gccagatctt aaaggagcat cctgtggaaa atacccaatt gcagaacgag

3061 aaactttacc tctattacct acaaaatgga agggacatgt atgttgatca ggaactggac

3121 ataaaccgtt tatctgatta cgacgtcgat gccattgtac cccaatcctt tttgaaggac

3181 gattcaatcg acaataaagt gcttacacgc tcggataaga accgagggaa aagtgacaat

3241 gttccaagcg aggaagtcgt aaagaaaatg aagaactatt ggcggcagct cctaaatgcg

3301 aaactgataa cgcaaagaaa gttcgataac ttaactaaag ctgagagggg tggcttgtct

3361 gaacttgaca aggccggatt tattaaacgt cagctcgtgg aaacccgcca aatcacaaag

3421 catgttgcac agatactaga ttcccgaatg aatacgaaat acgacgagaa cgataagctg

3481 attcgggaag tcaaagtaat cactttaaag tcaaaattgg tgtcggactt cagaaaggat

3541 tttcaattct ataaagttag ggagataaat aactaccacc atgcgcacga cgcttatctt

3601 aatgccgtcg tagggaccgc actcattaag aaatacccga agctagaaag tgagtttgtg

3661 tatggtgatt acaaagttta tgacgtccgt aagatgatcg cgaaaagcga acaggagata

3721 ggcaaggcta cagccaaata cttcttttat tctaacatta tgaatttctt taagacggaa

3781 atcactctgg caaacggaga gatacgcaaa cgacctttaa ttgaaaccaa tggggagaca

3841 ggtgaaatcg tatgggataa gggccgggac ttcgcgacgg tgagaaaagt tttgtccatg

3901 ccccaagtca acatagtaaa gaaaactgag gtgcagaccg gagggttttc aaaggaatcg

3961 attcttccaa aaaggaatag tgataagctc atcgctcgta aaaaggactg ggacccgaaa

4021 aagtacggtg gcttcgatag ccctacagtt gcctattctg tcctagtagt ggcaaaagtt

4081 gagaagggaa aatccaagaa actgaagtca gtcaaagaat tattggggat aacgattatg

4141 gagcgctcgt cttttgaaaa gaaccccatc gacttccttg aggcgaaagg ttacaaggaa

4201 gtaaaaaagg atctcataat taaactacca aagtatagtc tgtttgagtt agaaaatggc

4261 cgaaaacgga tgttggctag cgccggagag cttcaaaagg ggaacgaact cgcactaccg

4321 tctaaatacg tgaatttcct gtatttagcg tcccattacg agaagttgaa aggttcacct

4381 gaagataacg aacagaagca actttttgtt gagcagcaca aacattatct cgacgaaatc

4441 atagagcaaa tttcggaatt cagtaagaga gtcatcctag ctgatgccaa tctggacaaa

4501 gtattaagcg catacaacaa gcacagggat aaacccatac gtgagcaggc ggaaaatatt

4561 atccatttgt ttactcttac caacctcggc gctccagccg cattcaagta ttttgacaca

4621 acgatagatc gcaaacgata cacttctacc aaggaggtgc tagacgcgac actgattcac

4681 caatccatca cgggattata tgaaactcgg atagatttgt cacagcttgg gggtgacgga

4741 tcccccaaga agaagaggaa agtctga.

In certain embodiments, the dCas9 of the disclosure comprises a dCas9 isolated or derived from Staphyloccocus pyogenes. In certain embodiments, the dCas9 comprises a dCas9 with substitutions at positions 10 and 840 of the amino acid sequence of the dCas9, which inactivate the catalytic site. In certain embodiments, these substitutions are D10A and H840A. In certain embodiments, the “X” residue at position 1 of the dCas9 sequence is a methionine (M). In certain embodiments, the amino acid sequence of the dCas9 comprises the sequence of:

(SEQ ID NO: 14498)

1 XDKKYSIGLA IGTNSVGWAV ITDEYKVPSK KFKVLGNTDR HSIKKNLIGA LLFDSGETAE

61 ATRLKRTARR RYTRRKNRIC YLQEIFSNEM AKVDDSFFHR LEESFLVEED KKHERHPIFG

121 NIVDEVAYHE KYPTIYHLRK KLVDSTDKAD LRLIYLALAH MIKFRGHFLI EGDLNPDNSD

181 VDKLFIQLVQ TYNQLFEENP INASGVDAKA ILSARLSKSR RLENLIAQLP GEKKNGLFGN

241 LIALSLGLTP NFKSNFDLAE DAKLQLSKDT YDDDLDNLLA QIGDQYADLF LAAKNLSDAI

301 LLSDILRVNT EITKAPLSAS MIKRYDEHHQ DLTLLKALVR QQLPEKYKEI FFDQSKNGYA

361 GYIDGGASQE EFYKFIKPIL EKMDGTEELL VKLNREDLLR KQRTFDNGSI PHQIHLGELH

421 AILRRQEDFY PFLKDNREKI EKILTFRIPY YVGPLARGNS RFAWMTRKSE ETITPWNFEE

481 VVDKGASAQS FIERMTNFDK NLPNEKVLPK HSLLYEYFTV YNELTKVKYV TEGMRKPAFL

541 SGEQKKAIVD LLFKTNRKVT VKQLKEDYFK KIECFDSVEI SGVEDRFNAS LGTYHDLLKI

601 IKDKDFLDNE ENEDILEDIV LTLTLFEDRE MIEERLKTYA HLFDDKVMKQ LKRRRYTGWG

661 RLSRKLINGI RDKQSGKTIL DFLKSDGFAN RNFMQLIHDD SLTFKEDIQK AQVSGQGDSL

721 HEHIANLAGS PAIKKGILQT VKVVDELVKV MGRHKPENIV IEMARENQTT QKGQKNSRER

781 MKRIEEGIKE LGSQILKEHP VENTQLQNEK LYLYYLQNGR DMYVDQELDI NRLSDYDVDA

841 IVPQSFLKDD SIDNKVLTRS DKNRGKSDNV PSEEVVKKMK NYWRQLLNAK LITQRKFDNL

901 TKAERGGLSE LDKAGFIKRQ LVETRQITKH VAQILDSRMN TKYDENDKLI REVKVITLKS

961 KLVSDFRKDF QFYKVREINN YHHAHDAYLN AVVGTALIKK YPKLESEFVY GDYKVYDVRK

1021 MIAKSEQEIG KATAKYFFYS NIMNFFKTEI TLANGEIRKR PLIETNGETG EIVWDKGRDF

1081 ATVRKVLSMP QVNIVKKTEV QTGGFSKESI LPKRNSDKLI ARKKDWDPKK YGGFDSPTVA

1141 YSVLVVAKVE KGKSKKLKSV KELLGITIME RSSFEKNPID FLEAKGYKEV KKDLIIKLPK

1201 YSLFELENGR KRMLASAGEL QKGNELALPS KYVNFLYLAS HYEKLKGSPE DNEQKQLFVE

1261 QHKHYLDEII EQISEFSKRV ILADANLDKV LSAYNKHRDK PIREQAENII HLFTLTNLGA

1321 PAAFKYFDTT IDRKRYTSTK EVLDATLIHQ SITGLYETRI DLSQLGGD.

In certain embodiments, the dCas9 of the disclosure comprises a dCas9 isolated or derived from Staphylococcus aureus. In certain embodiments, the dCas9 comprises a dCas9 with substitutions at positions 10 and 580 of the amino acid sequence of the dCas9 which inactivate the catalytic site. In certain embodiments, these substitutions are D10A and N580A. In certain embodiments, the dCas9 is a small and inactive Cas9 (dSaCas9). In certain embodiments, the amino acid sequence of the dSaCas9 comprises the sequence of:

(SEQ ID NO: 14658)

1 mkrnyilglA igitsvgygi idyetrdvid agvrlfkean vennegrrsk rgarrlkrrr

61 rhriqrvkkl lfdynlltdh selsginpye arvkglsqkl seeefsaall hlakrrgvhn

121 vneveedtgn elstkeqisr nskaleekyv aelqlerlkk dgevrgsinr fktsdyvkea

181 kqllkvqkay hqldqsfidt yidlletrrt yyegpgegsp fgwkdikewy emlmghctyf

241 peelrsvkya ynadlynaln dlnnlvitrd enekleyyek fqiienvfkq kkkptlkqia

301 keilvneedi kgyrvtstgk peftnlkvyh dikditarke iienaelldq iakiltiyqs

361 sediqeeltn lnseltqeei eqisnlkgyt gthnlslkai nlildelwht ndnqiaifnr

421 lklvpkkvd1 sqqkeipttl vddfilspvv krsfiqsikv inaiikkygl pndiiielar

481 eknskdaqkm inemqkrnrq tnerieeiir ttgkenakyl iekiklhdmq egkclyslea

541 ipledllnnp fnyevdhiip rsysfdnsfn nkvlvkqeeA skkgnrtpfq ylsssdskis

601 yetfkkhiln lakgkgrisk tkkeylleer dinrfsvqkd finrnlvdtr yatrglmnll

661 rsyfrvnnld vkvksinggf tsflrrkwkf kkernkgykh haedaliian adfifkewkk

721 ldkakkvmen qmfeekqaes mpeieteqey keifitphqi khikdfkdyk yshrvdkkpn

781 relindtlys trkddkgntl ivnnlnglyd kdndklkkli nkspekllmy hhdpqtyqkl

841 klimeqygde knplykyyee tgnyltkysk kdngpvikki kyygnklnah lditddypns

901 rnkvvklslk pyrfdvyldn gvykfvtvkn ldvikkenyy evnskcyeea kklkkisnqa

961 efiasfynnd likingelyr vigvnndlln rievnmidit yreylenmnd krppriikti

1021 asktqsikky stdilgnlye vkskkhpqii kkg.

In certain embodiments of the gene editing systems described herein, the nuclease may comprise, consist essentially of or consist of, a homodimer or a heterodimer. Nuclease domains of the disclosure may comprise, consist essentially of or consist of a nuclease domain isolated, derived or recombined from a transcription-activator-like effector nuclease (TALEN). TALENs are transcription factors with programmable DNA binding domains that provide a means to create designer proteins that bind to pre-determined DNA sequences or individual nucleic acids. Modular DNA binding domains have been identified in transcriptional activator-like (TAL) proteins, or, more specifically, transcriptional activator-like effector nucleases (TALENs), thereby allowing for the de novo creation of synthetic transcription factors that bind to DNA sequences of interest and, if desirable, also allowing a second domain present on the protein or polypeptide to perform an activity related to DNA. TAL proteins have been derived from the organisms Xanthomonas and Ralstonia.

In certain embodiments of the gene editing systems described herein, the nuclease domain may comprise, consist essentially of or consist of a nuclease domain isolated, derived or recombined from a TALEN and a type IIS endonuclease. In certain embodiments of the disclosure, the type IIS endonuclease may comprise, consist essentially of or consist of AciI, Mn1I, AlwI, BbvI, BccI, BceAI, BsmAI, BsmFI, BspCNI, BsrI, BtsCI, HgaI, HphI, HpyAV, MbolI, My1I, PleI, SfaNI, AcuI, BciVI, BfuAI, BmgBI, BmrI, BpmI, BpuEI, BsaI, BseRI, BsgI, BsmI, BspMI, BsrBI, BsrBI, BsrDI, BtgZI, BtsI, EarI, EciI, MmeI, NmeAIII, BbvCI, Bpu10I, BspQI, SapI, BaeI, BsaXI, CspCI, BfiI, MboII, Acc36I, FokI or Clo051. In certain embodiments of the disclosure, the type IIS endonuclease may comprise, consist essentially of or consist of Clo051 (SEQ ID NO: 14503).

In certain embodiments of the gene editing systems described herein, the nuclease domain of may comprise, consist essentially of or consist of a nuclease domain isolated, derived or recombined from a zinc finger nuclease (ZFN) and a type IIS endonuclease. In certain embodiments of the disclosure, the type IIS endonuclease may comprise, consist essentially of or consist of AciI, Mn1I, AlwI, BbvI, BccI, BceAI, BsmAI, BsmFI, BspCNI, BsrI, BtsCI, HgaI, HphI, HpyAV, Mbo1I, My1I, PleI, SfaNI, AcuI, BciVI, BfuAI, BmgBI, BmrI, BpmI, BpuEI, BsaI, BseRI, BsgI, BsmI, BspMI, BsrBI, BsrBI, BsrDI, BtgZI, BtsI, EarI, EciI, MmeI, NmeAIII, BbvCI, Bpu10I, BspQI, SapI, BaeI, BsaXI, CspCI, BfiI, MboII, Acc36I, FokI or Clo051. In certain embodiments of the disclosure, the type IIS endonuclease may comprise, consist essentially of or consist of Clo051 (SEQ ID NO: 14503).

In certain embodiments of the gene editing systems described herein, the DNA binding domain and the nuclease domain may be covalently linked. For example, a fusion protein may comprise the DNA binding domain and the nuclease domain. In certain embodiments of the genomic editing compositions or constructs of the disclosure, the DNA binding domain and the nuclease domain may be operably linked through a non-covalent linkage.

Therapeutic Proteins

In certain embodiments of the composition and methods of the disclosure, modified immune or immune precursor cells express therapeutic proteins. Therapeutic proteins of the disclosure include secreted proteins. Preferably, in a therapeutic context, the therapeutic protein is a human protein, including a secreted human protein. When expressed or secreted by immune or immune precursor cells of the disclosure, the combination comprising the immune or immune precursor cell and the therapeutic protein secreted therefrom may be considered a monotherapy. However, the immune or immune precursor cells of the disclosure may be administered as a combination therapy with a second agent. Human therapeutic proteins of the disclosure include, but are not limited to, those provided at Table 1.

TABLE 1

Exemplary Human Secreted Proteins

Gene Name
Gene Description
Protein SEQ ID NO

A1BG
Alpha-1-B glycoprotein
SEQ ID NOS: 1-2

A2M
Alpha-2-macroglobulin
SEQ ID NOS: 3-6

A2ML1
Alpha-2-macroglobulin-like 1
SEQ ID NOS: 7-12

A4GNT
Alpha-1,4-N-acetylglucosaminyltransferase
SEQ ID NO: 13

AADACL2
Arylacetamide deacetylase-like 2
SEQ ID NOS: 14-15

AANAT
Aralkylamine N-acetyltransferase
SEQ ID NOS: 16-19

ABCG1
ATP-binding cassette, sub-family G (WHITE),
SEQ ID NOS: 20-26

member 1

ABHD1
Abhydrolase domain containing 1
SEQ ID NOS: 27-31

ABHD10
Abhydrolase domain containing 10
SEQ ID NOS: 32-35

ABHD14A
Abhydrolase domain containing 14A
SEQ ID NOS: 36-40

ABHD15
Abhydrolase domain containing 15
SEQ ID NO: 41

ABI3BP
ABI family, member 3 (NESH) binding protein
SEQ ID NOS: 42-63

FAM175A
Family with sequence similarity 175, member A
SEQ ID NOS: 64-71

LA16c-

SEQ ID NO: 72

380H5.3

AC008641.1

SEQ ID NO: 73

CTB-

SEQ ID NOS: 74-75

601318.6

AC009133.22

SEQ ID NO: 76

AC009491.2

SEQ ID NO: 77

RP11-

SEQ ID NOS: 78-80

977G19.10

CTD-

SEQ ID NOS: 81-84

2370N5.3

RP11-

SEQ ID NOS: 85-87

196G11.1

AC136352.5

SEQ ID NO: 88

RP11-

SEQ ID NO: 89

812E19.9

AC145212.4
MaFF-interacting protein
SEQ ID NO: 90

AC233755.1

SEQ ID NO: 91

AC011513.3

SEQ ID NOS: 92-93

ACACB
Acetyl-CoA carboxylase beta
SEQ ID NOS: 94-100

ACAN
Aggrecan
SEQ ID NOS: 101-108

ACE
Angiotensin I converting enzyme
SEQ ID NOS: 109-121

ACHE
Acetylcholinesterase (Yt blood group)
SEQ ID NOS: 122-134

ACP2
Acid phosphatase 2, lysosomal
SEQ ID NOS: 135-142

ACP5
Acid phosphatase 5, tartrate resistant
SEQ ID NOS: 143-151

ACP6
Acid phosphatase 6, lysophosphatidic
SEQ ID NOS: 152-158

PAPL
Iron/zinc purple acid phosphatase-like protein
SEQ ID NOS: 159-162

ACPP
Acid phosphatase, prostate
SEQ ID NOS: 163-167

ACR
Acrosin
SEQ ID NOS: 168-169

ACRBP
Acrosin binding protein
SEQ ID NOS: 170-174

ACRV1
Acrosomal vesicle protein 1
SEQ ID NOS: 175-178

ACSF2
Acyl-CoA synthetase family member 2
SEQ ID NOS: 179-187

ACTL10
Actin-like 10
SEQ ID NO: 188

ACVR1
Activin A receptor, type I
SEQ ID NOS: 189-197

ACVR1C
Activin A receptor, type IC
SEQ ID NOS: 198-201

ACVRL1
Activin A receptor type II-like 1
SEQ ID NOS: 202-207

ACYP1
Acylphosphatase 1, erythrocyte (common) type
SEQ ID NOS: 208-213

ACYP2
Acylphosphatase 2, muscle type
SEQ ID NOS: 214-221

CECR1
Cat eye syndrome chromosome region, candidate 1
SEQ ID NOS: 222-229

ADAM10
ADAM metallopeptidase domain 10
SEQ ID NOS: 230-237

ADAM12
ADAM metallopeptidase domain 12
SEQ ID NOS: 238-240

ADAM15
ADAM metallopeptidase domain 15
SEQ ID NOS: 241-252

ADAM17
ADAM metallopeptidase domain 17
SEQ ID NOS: 253-255

ADAM18
ADAM metallopeptidase domain 18
SEQ ID NOS: 256-260

ADAM22
ADAM metallopeptidase domain 22
SEQ ID NOS: 261-269

ADAM28
ADAM metallopeptidase domain 28
SEQ ID NOS: 270-275

ADAM29
ADAM metallopeptidase domain 29
SEQ ID NOS: 276-284

ADAM32
ADAM metallopeptidase domain 32
SEQ ID NOS: 285-291

ADAM33
ADAM metallopeptidase domain 33
SEQ ID NOS: 292-296

ADAM7
ADAM metallopeptidase domain 7
SEQ ID NOS: 297-300

ADAM8
ADAM metallopeptidase domain 8
SEQ ID NOS: 301-305

ADAM9
ADAM metallopeptidase domain 9
SEQ ID NOS: 306-311

ADAMDEC1
ADAM-like, decysin 1
SEQ ID NOS: 312-314

ADAMTS1
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 315-318

1 motif, 1

ADAMTS10
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 319-324

1 motif, 10

ADAMTS12
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 325-327

1 motif, 12

ADAMTS13
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 328-335

1 motif, 13

ADAMTS14
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 336-337

1 motif, 14

ADAMTS15
ADAM metallopeptidase with thrombospondin type
SEQ ID NO: 338

1 motif, 15

ADAMTS16
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 339-340

1 motif, 16

ADAMTS17
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 341-344

1 motif, 17

ADAMTS18
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 345-348

1 motif, 18

ADAMTS19
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 349-352

1 motif, 19

ADAMTS2
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 353-355

1 motif, 2

ADAMTS20
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 356-359

1 motif, 20

ADAMTS3
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 360-361

1 motif, 3

ADAMTS5
ADAM metallopeptidase with thrombospondin type
SEQ ID NO: 362

1 motif, 5

ADAMTS6
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 363-364

1 motif, 6

ADAMTS7
ADAM metallopeptidase with thrombospondin type
SEQ ID NO: 365

1 motif, 7

ADAMTS8
ADAM metallopeptidase with thrombospondin type
SEQ ID NO: 366

1 motif, 8

ADAMTS9
ADAM metallopeptidase with thrombospondin type
SEQ ID NOS: 367-371

1 motif, 9

ADAMTSL1
ADAMTS-like 1
SEQ ID NOS: 372-382

ADAMTSL2
ADAMTS-like 2
SEQ ID NOS: 383-385

ADAMTSL3
ADAMTS-like 3
SEQ ID NOS: 386-387

ADAMTSL4
ADAMTS-like 4
SEQ ID NOS: 388-391

ADAMTSL5
ADAMTS-like 5
SEQ ID NOS: 392-397

ADCK1
AarF domain containing kinase 1
SEQ ID NOS: 398-402

ADCYAP1
Adenylate cyclase activating polypeptide 1
SEQ ID NOS: 403-404

(pituitary)

ADCYAP1R1
Adenylate cyclase activating polypeptide 1
SEQ ID NOS: 405-411

(pituitary) receptor type I

ADGRA3
Adhesion G protein-coupled receptor A3
SEQ ID NOS: 412-416

ADGRB2
Adhesion G protein-coupled receptor B2
SEQ ID NOS: 417-425

ADGRD1
Adhesion G protein-coupled receptor D1
SEQ ID NOS: 426-431

ADGRE3
Adhesion G protein-coupled receptor E3
SEQ ID NOS: 432-436

ADGRE5
Adhesion G protein-coupled receptor E5
SEQ ID NOS: 437-442

ADGRF1
Adhesion G protein-coupled receptor F1
SEQ ID NOS: 443-447

ADGRG1
Adhesion G protein-coupled receptor G1
SEQ ID NOS: 448-512

ADGRG5
Adhesion G protein-coupled receptor G5
SEQ ID NOS: 513-515

ADGRG6
Adhesion G protein-coupled receptor G6
SEQ ID NOS: 516-523

ADGRV1
Adhesion G protein-coupled receptor V1
SEQ ID NOS: 524-540

ADI1
Acireductone dioxygenase 1
SEQ ID NOS: 541-543

ADIG
Adipogenin
SEQ ID NOS: 544-547

ADIPOQ
Adiponectin, C1Q and collagen domain containing
SEQ ID NOS: 548-549

ADM
Adrenomedullin
SEQ ID NOS: 550-557

ADM2
Adrenomedullin 2
SEQ ID NOS: 558-559

ADM5
Adrenomedullin 5 (putative)
SEQ ID NO: 560

ADPGK
ADP-dependent glucokinase
SEQ ID NOS: 561-570

ADPRHL2
ADP-ribosylhydrolase like 2
SEQ ID NO: 571

AEBP1
AE binding protein 1
SEQ ID NOS: 572-579

LACE1
Lactation elevated 1
SEQ ID NOS: 580-583

AFM
Afamin
SEQ ID NO: 584

AFP
Alpha-fetoprotein
SEQ ID NOS: 585-586

AGA
Aspartylglucosaminidase
SEQ ID NOS: 587-589

AGER
Advanced glycosylation end product-specific
SEQ ID NOS: 590-600

receptor

AGK
Acylglycerol kinase
SEQ ID NOS: 601-606

AGPS
Alkylglycerone phosphate synthase
SEQ ID NOS: 607-610

AGR2
Anterior gradient 2, protein disulphide isomerase
SEQ ID NOS: 611-614

family member

AGR3
Anterior gradient 3, protein disulphide isomerase
SEQ ID NOS: 615-617

family member

AGRN
Agrin
SEQ ID NOS: 618-621

AGRP
Agouti related neuropeptide
SEQ ID NO: 622

AGT
Angiotensinogen (serpin peptidase inhibitor, clade A,
SEQ ID NO: 623

member 8)

AGTPBP1
ATP/GTP binding protein 1
SEQ ID NOS: 624-627

AGTRAP
Angiotensin 11 receptor-associated protein
SEQ ID NOS: 628-635

AHCYL2
Adenosylhomocysteinase-like 2
SEQ ID NOS: 636-642

AHSG
Alpha-2-HS-glycoprotein
SEQ ID NOS: 643-644

AIG1
Androgen-induced 1
SEQ ID NOS: 645-653

AK4
Adenylate kinase 4
SEQ ID NOS: 654-657

AKAP10
A kinase (PRKA) anchor protein 10
SEQ ID NOS: 658-666

AKR1C1
Aldo-keto reductase family 1, member C1
SEQ ID NOS: 667-669

RP4-

SEQ ID NOS: 670-672

576H24.4

SERPINA3
Serpin peptidase inhibitor, clade A (alpha-1
SEQ ID NO: 673

antiproteinase, antitrypsin), member 3

RP11-14J7.7

SEQ ID NOS: 674-675

RP11-

SEQ ID NO: 676

903H12.5

AL356289.1

SEQ ID NO: 677

AL589743.1

SEQ ID NO: 678

XXbac-

SEQ ID NOS: 679-680

BPG116M5.17

XXbac-

SEQ ID NO: 681

BPG181M17.5

XXbac-

SEQ ID NO: 682

BPG32J3.20

RP11-

SEQ ID NO: 683

350O14.18

ALAS2
5′-aminolevulinate synthase 2
SEQ ID NOS: 684-691

ALB
Albumin
SEQ ID NOS: 692-701

ALDH9A1
Aldehyde dehydrogenase 9 family, member A1
SEQ ID NO: 702

ALDOA
Aldolase A, fructose-bisphosphate
SEQ ID NOS: 703-717

ALG1
ALG1, chitobiosyldiphosphodolichol beta-
SEQ ID NOS: 718-723

mannosyltransferase

ALG5
ALG5, dolichyl-phosphate beta-glucosyltransferase
SEQ ID NOS: 724-725

ALG9
ALG9, alpha-1,2-mannosyltransferase
SEQ ID NOS: 726-736

FAM150A
Family with sequence similarity 150, member A
SEQ ID NOS: 737-738

FAM150B
Family with sequence similarity 150, member B
SEQ ID NOS: 739-745

ALKBH1
AlkB homolog 1, histone H2A dioxygenase
SEQ ID NOS: 746-748

ALKBH5
AlkB homolog 5, RNA demethylase
SEQ ID NOS: 749-750

ALPI
Alkaline phosphatase, intestinal
SEQ ID NOS: 751-752

ALPL
Alkaline phosphatase, liver/bone/kidney
SEQ ID NOS: 753-757

ALPP
Alkaline phosphatase, placental
SEQ ID NO: 758

ALPPL2
Alkaline phosphatase, placental-like 2
SEQ ID NO: 759

AMBN
Ameloblastin (enamel matrix protein)
SEQ ID NOS: 760-762

AMBP
Alpha-1-microglobulin/bikunin precursor
SEQ ID NOS: 763-765

AMELX
Amelogenin, X-linked
SEQ ID NOS: 766-768

AMELY
Amelogenin, Y-linked
SEQ ID NOS: 769-770

AMH
Anti-Mullerian hormone
SEQ ID NO: 771

AMPD1
Adenosine monophosphate deaminase 1
SEQ ID NOS: 772-774

AMTN
Amelotin
SEQ ID NOS: 775-776

AMY1A
Amylase, alpha 1A (salivary)
SEQ ID NOS: 777-779

AMY1B
Amylase, alpha 1B (salivary)
SEQ ID NOS: 780-783

AMY1C
Amylase, alpha 1C (salivary)
SEQ ID NO: 784

AMY2A
Amylase, alpha 2A (pancreatic)
SEQ ID NOS: 785-787

AMY2B
Amylase, alpha 2B (pancreatic)
SEQ ID NOS: 788-792

ANG
Angiogenin, ribonuclease, RNase A family, 5
SEQ ID NOS: 793-794

ANGEL1
Angel homolog 1 (Drosophila)
SEQ ID NOS: 795-798

ANGPT1
Angiopoietin 1
SEQ ID NOS: 799-803

ANGPT2
Angiopoietin 2
SEQ ID NOS: 804-807

ANGPT4
Angiopoietin 4
SEQ ID NO: 808

ANGPTL1
Angiopoietin-like 1
SEQ ID NOS: 809-811

ANGPTL2
Angiopoietin-like 2
SEQ ID NOS: 812-813

ANGPTL3
Angiopoietin-like 3
SEQ ID NO: 814

ANGPTL4
Angiopoietin-like 4
SEQ ID NOS: 815-822

ANGPTL5
Angiopoietin-like 5
SEQ ID NOS: 823-824

ANGPTL6
Angiopoietin-like 6
SEQ ID NOS: 825-827

ANGPTL7
Angiopoietin-like 7
SEQ ID NO: 828

C19orf80
Chromosome 19 open reading frame 80
SEQ ID NOS: 829-832

ANK1
Ankyrin 1, erythrocytic
SEQ ID NOS: 833-843

ANKDD1A
Ankyrin repeat and death domain containing 1A
SEQ ID NOS: 844-850

ANKRD54
Ankyrin repeat domain 54
SEQ ID NOS: 851-859

ANKRD60
Ankyrin repeat domain 60
SEQ ID NO: 860

ANO7
Anoctamin 7
SEQ ID NOS: 861-864

ANOS1
Anosmin 1
SEQ ID NO: 865

ANTXR1
Anthrax toxin receptor 1
SEQ ID NOS: 866-869

AOAH
Acyloxyacyl hydrolase (neutrophil)
SEQ ID NOS: 870-874

AOC1
Amine oxidase, copper containing 1
SEQ ID NOS: 875-880

AOC2
Amine oxidase, copper containing 2 (retina-specific)
SEQ ID NOS: 881-882

AOC3
Amine oxidase, copper containing 3
SEQ ID NOS: 883-889

AP000721.4

SEQ ID NO: 890

APBB1
Amyloid beta (A4) precursor protein-binding, family
SEQ ID NOS: 891-907

B, member 1 (Fe65)

APCDD1
Adenomatosis polyposis coli down-regulated 1
SEQ ID NOS: 908-913

APCS
Amyloid P component, serum
SEQ ID NO: 914

APELA
Apelin receptor early endogenous ligand
SEQ ID NOS: 915-917

APLN
Apelin
SEQ ID NO: 918

APLP2
Amyloid beta (A4) precursor-like protein 2
SEQ ID NOS: 919-928

APOA1
Apolipoprotein A-I
SEQ ID NOS: 929-933

APOA2
Apolipoprotein A-II
SEQ ID NOS: 934-942

APOA4
Apolipoprotein A-IV
SEQ ID NO: 943

APOA5
Apolipoprotein A-V
SEQ ID NOS: 944-946

APOB
Apolipoprotein B
SEQ ID NOS: 947-948

APOC1
Apolipoprotein C-I
SEQ ID NOS: 949-957

APOC2
Apolipoprotein C-II
SEQ ID NOS: 958-962

APOC3
Apolipoprotein C-III
SEQ ID NOS: 963-966

APOC4
Apolipoprotein C-IV
SEQ ID NOS: 967-968

APOC4-
APOC4-APOC2 readthrough (NMD candidate)
SEQ ID NOS: 969-970

APOC2

APOD
Apolipoprotein D
SEQ ID NOS: 971-974

APOE
Apolipoprotein E
SEQ ID NOS: 975-978

APOF
Apolipoprotein F
SEQ ID NO: 979

APOH
Apolipoprotein H (beta-2-glycoprotein I)
SEQ ID NOS: 980-983

APOL1
Apolipoprotein L, 1
SEQ ID NOS: 984-994

APOL3
Apolipoprotein L, 3
SEQ ID NOS: 995-1009

APOM
Apolipoprotein M
SEQ ID NOS: 1010-1012

APOOL
Apolipoprotein O-like
SEQ ID NOS: 1013-1015

ARCN1
Archain 1
SEQ ID NOS: 1016-1020

ARFIP2
ADP-ribosylation factor interacting protein 2
SEQ ID NOS: 1021-1027

ARHGAP36
Rho GTPase activating protein 36
SEQ ID NOS: 1028-1033

HMHA1
Histocompatibility (minor) HA-1
SEQ ID NOS: 1034-1042

ARHGAP6
Rho GTPase activating protein 6
SEQ ID NOS: 1043-1048

ARHGEF4
Rho guanine nucleotide exchange factor (GEF) 4
SEQ ID NOS: 1049-1059

ARL16
ADP-ribosylation factor-like 16
SEQ ID NOS: 1060-1068

ARMC5
Armadillo repeat containing 5
SEQ ID NOS: 1069-1075

ARNTL
Aryl hydrocarbon receptor nuclear translocator-like
SEQ ID NOS: 1076-1090

ARSA
Arylsulfatase A
SEQ ID NOS: 1091-1096

ARSB
Arylsulfatase B
SEQ ID NOS: 1097-1100

ARSE
Arylsulfatase E (chondrodysplasia punctata 1)
SEQ ID NOS: 1101-1104

ARSG
Arylsulfatase G
SEQ ID NOS: 1105-1108

ARSI
Arylsulfatase family, member I
SEQ ID NOS: 1109-1111

ARSK
Arylsulfatase family, member K
SEQ ID NOS: 1112-1116

ART3
ADP-ribosyltransferase 3
SEQ ID NOS: 1117-1124

ART4
ADP-ribosyltransferase 4 (Dombrock blood group)
SEQ ID NOS: 1125-1128

ART5
ADP-ribosyltransferase 5
SEQ ID NOS: 1129-1133

ARTN
Artemin
SEQ ID NOS: 1134-1144

ASAH1
N-acylsphingosine amidohydrolase (acid
SEQ ID NOS: 1145-1195

ceramidase) 1

ASAH2
N-acylsphingosine amidohydrolase (non-lysosomal
SEQ ID NOS: 1196-1201

ceramidase) 2

ASCL1
Achaete-scute family bHLH transcription factor 1
SEQ ID NO: 1202

ASIP
Agouti signaling protein
SEQ ID NOS: 1203-1204

ASPN
Asporin
SEQ ID NOS: 1205-1206

ASTL
Astacin-like metallo-endopeptidase (M12 family)
SEQ ID NO: 1207

ATAD5
ATPase family, AAA domain containing 5
SEQ ID NOS: 1208-1209

ATAT1
Alpha tubulin acetyltransferase 1
SEQ ID NOS: 1210-1215

ATG2A
Autophagy related 2A
SEQ ID NOS: 1216-1218

ATG5
Autophagy related 5
SEQ ID NOS: 1219-1227

ATMIN
ATM interactor
SEQ ID NOS: 1228-1231

ATP13A1
ATPase type 13A1
SEQ ID NOS: 1232-1234

ATP5F1
ATP synthase, H+ transporting, mitochondrial Fo
SEQ ID NOS: 1235-1236

complex, subunit B1

ATP6AP1
ATPase, H+ transporting, lysosomal accessory
SEQ ID NOS: 1237-1244

protein 1

ATP6AP2
ATPase, H+ transporting, lysosomal accessory
SEQ ID NOS: 1245-1267

protein 2

ATPAF1
ATP synthase mitochondrial F1 complex assembly
SEQ ID NOS: 1268-1278

factor 1

AUH
AU RNA binding protein/enoyl-CoA hydratase
SEQ ID NOS: 1279-1280

AVP
Arginine vasopressin
SEQ ID NO: 1281

AXIN2
Axin 2
SEQ ID NOS: 1282-1289

AZGP1
Alpha-2-glycoprotein 1, zinc-binding
SEQ ID NOS: 1290-1292

AZU1
Azurocidin 1
SEQ ID NOS: 1293-1294

B2M
Beta-2-microglobulin
SEQ ID NOS: 1295-1301

B3GALNT1
Beta-1,3-N-acetylgalactosaminyltransferase 1
SEQ ID NOS: 1302-1314

(globoside blood group)

B3GALNT2
Beta-1,3-N-acetylgalactosaminyltransferase 2
SEQ ID NOS: 1315-1317

B3GALT1
UDP-Gal:betaGlcNAc beta 1,3-galactosyltransferase,
SEQ ID NO: 1318

polypeptide 1

B3GALT4
UDP-Gal:betaGlcNAc beta 1,3-galactosyltransferase,
SEQ ID NO: 1319

polypeptide 4

B3GALT5
UDP-Gal:betaGlcNAc beta 1,3-galactosyltransferase,
SEQ ID NOS: 1320-1324

polypeptide 5

B3GALT6
UDP-Gal:betaGal beta 1,3-galactosyltransferase
SEQ ID NO: 1325

polypeptide 6

B3GAT3
Beta-1,3-glucuronyltransferase 3
SEQ ID NOS: 1326-1330

B3GLCT
Beta 3-glucosyltransferase
SEQ ID NO: 1331

B3GNT3
UDP-GlcNAc:betaGal beta-1,3-N-
SEQ ID NOS: 1332-1335

acetylglucosaminyltransferase 3

B3GNT4
UDP-GlcNAc:betaGal beta-1,3-N-
SEQ ID NOS: 1336-1339

acetylglucosaminyltransferase 4

B3GNT6
UDP-GlcNAc:betaGal beta-1,3-N-
SEQ ID NOS: 1340-1341

acetylglucosaminyltransferase 6

B3GNT7
UDP-GlcNAc:betaGal beta-1,3-N-
SEQ ID NO: 1342

acetylglucosaminyltransferase 7

B3GNT8
UDP-GlcNAc:betaGal beta-1,3-N-
SEQ ID NO: 1343

acetylglucosaminyltransferase 8

B3GNT9
UDP-GlcNAc:betaGal beta-1,3-N-
SEQ ID NO: 1344

acetylglucosaminyltransferase 9

B4GALNT1
Beta-1,4-N-acetyl-galactosaminyl transferase 1
SEQ ID NOS: 1345-1356

B4GALNT3
Beta-1,4-N-acetyl-galactosaminyl transferase 3
SEQ ID NOS: 1357-1358

B4GALNT4
Beta-1,4-N-acetyl-galactosaminyl transferase 4
SEQ ID NOS: 1359-1361

B4GALT4
UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase,
SEQ ID NOS: 1362-1375

polypeptide 4

B4GALT5
UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase,
SEQ ID NO: 1376

polypeptide 5

B4GALT6
UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase,
SEQ ID NOS: 1377-1380

polypeptide 6

B4GAT1
Beta-1,4-glucuronyltransferase 1
SEQ ID NO: 1381

B9D1
B9 protein domain 1
SEQ ID NOS: 1382-1398

BACE2
Beta-site APP-cleaving enzyme 2
SEQ ID NOS: 1399-1401

BAGE5
B melanoma antigen family, member 5
SEQ ID NO: 1402

BCAM
Basal cell adhesion molecule (Lutheran blood group)
SEQ ID NOS: 1403-1406

BCAN
Brevican
SEQ ID NOS: 1407-1413

BCAP29
B-cell receptor-associated protein 29
SEQ ID NOS: 1414-1426

BCAR1
Breast cancer anti-estrogen resistance 1
SEQ ID NOS: 1427-1444

BCHE
Butyrylcholinesterase
SEQ ID NOS: 1445-1449

BCKDHB
Branched chain keto acid dehydrogenase E1, beta
SEQ ID NOS: 1450-1452

polypeptide

BDNF
Brain-derived neurotrophic factor
SEQ ID NOS: 1453-1470

BGLAP
Bone gamma-carboxyglutamate (gla) protein
SEQ ID NO: 1471

BGN
Biglycan
SEQ ID NOS: 1472-1473

BLVRB
Biliverdin reductase B
SEQ ID NOS: 1474-1478

BMP1
Bone morphogenetic protein 1
SEQ ID NOS: 1479-1490

BMP10
Bone morphogenetic protein 10
SEQ ID NO: 1491

BMP15
Bone morphogenetic protein 15
SEQ ID NO: 1492

BMP2
Bone morphogenetic protein 2
SEQ ID NO: 1493

BMP3
Bone morphogenetic protein 3
SEQ ID NO: 1494

BMP4
Bone morphogenetic protein 4
SEQ ID NOS: 1495-1502

BMP6
Bone morphogenetic protein 6
SEQ ID NO: 1503

BMP7
Bone morphogenetic protein 7
SEQ ID NOS: 1504-1507

BMP8A
Bone morphogenetic protein 8a
SEQ ID NO: 1508

BMP8B
Bone morphogenetic protein 8b
SEQ ID NO: 1509

BMPER
BMP binding endothelial regulator
SEQ ID NOS: 1510-1513

BNC1
Basonuclin 1
SEQ ID NOS: 1514-1515

BOC
BOC cell adhesion associated, oncogene regulated
SEQ ID NOS: 1516-1526

BOD1
Biorientation of chromosomes in cell division 1
SEQ ID NOS: 1527-1531

BOLA1
BolA family member 1
SEQ ID NOS: 1532-1534

BPI
Bactericidal/permeability-increasing protein
SEQ ID NOS: 1535-1538

BPIFA1
BPI fold containing family A, member 1
SEQ ID NOS: 1539-1542

BPIFA2
BPI fold containing family A, member 2
SEQ ID NOS: 1543-1544

BPIFA3
BPI fold containing family A, member 3
SEQ ID NOS: 1545-1546

BPIFB1
BPI fold containing family B, member 1
SEQ ID NOS: 1547-1548

BPIFB2
BPI fold containing family B, member 2
SEQ ID NO: 1549

BPIFB3
BPI fold containing family B, member 3
SEQ ID NO: 1550

BPIFB4
BPI fold containing family B, member 4
SEQ ID NOS: 1551-1552

BPIFB6
BPI fold containing family B, member 6
SEQ ID NOS: 1553-1554

BPIFC
BPI fold containing family C
SEQ ID NOS: 1555-1558

BRF1
BRF1, RNA polymerase III transcription initiation
SEQ ID NOS: 1559-1574

factor 90 kDa subunit

BRINP1
Bone morphogenetic protein/retinoic acid inducible
SEQ ID NOS: 1575-1576

neural-specific 1

BRINP2
Bone morphogenetic protein/retinoic acid inducible
SEQ ID NO: 1577

neural-specific 2

BRINP3
Bone morphogenetic protein/retinoic acid inducible
SEQ ID NOS: 1578-1580

neural-specific 3

BSG
Basigin (Ok blood group)
SEQ ID NOS: 1581-1591

BSPH1
Binder of sperm protein homolog 1
SEQ ID NO: 1592

BST1
Bone marrow stromal cell antigen 1
SEQ ID NOS: 1593-1597

BTBD17
BTB (POZ) domain containing 17
SEQ ID NO: 1598

BTD
Biotinidase
SEQ ID NOS: 1599-1608

BTN2A2
Butyrophilin, subfamily 2, member A2
SEQ ID NOS: 1609-1622

BTN3A1
Butyrophilin, subfamily 3, member A1
SEQ ID NOS: 1623-1629

BTN3A2
Butyrophilin, subfamily 3, member A2
SEQ ID NOS: 1630-1640

BTN3A3
Butyrophilin, subfamily 3, member A3
SEQ ID NOS: 1641-1649

RP4-
Complement factor H-related protein 2
SEQ ID NO: 1650

608O15.3

C10orf99
Chromosome 10 open reading frame 99
SEQ ID NO: 1651

C11orf1
Chromosome 11 open reading frame 1
SEQ ID NOS: 1652-1656

C11orf24
Chromosome 11 open reading frame 24
SEQ ID NOS: 1657-1659

C11orf45
Chromosome 11 open reading frame 45
SEQ ID NOS: 1660-1661

C11orf94
Chromosome 11 open reading frame 94
SEQ ID NO: 1662

C12orf10
Chromosome 12 open reading frame 10
SEQ ID NOS: 1663-1666

C12orf49
Chromosome 12 open reading frame 49
SEQ ID NOS: 1667-1670

C12orf73
Chromosome 12 open reading frame 73
SEQ ID NOS: 1671-1680

C12orf76
Chromosome 12 open reading frame 76
SEQ ID NOS: 1681-1688

C14orf93
Chromosome 14 open reading frame 93
SEQ ID NOS: 1689-1704

C16orf89
Chromosome 16 open reading frame 89
SEQ ID NOS: 1705-1707

C16orf90
Chromosome 16 open reading frame 90
SEQ ID NOS: 1708-1709

C17orf67
Chromosome 17 open reading frame 67
SEQ ID NO: 1710

C17orf75
Chromosome 17 open reading frame 75
SEQ ID NOS: 1711-1719

C17orf99
Chromosome 17 open reading frame 99
SEQ ID NOS: 1720-1722

C18orf54
Chromosome 18 open reading frame 54
SEQ ID NOS: 1723-1727

C19orf47
Chromosome 19 open reading frame 47
SEQ ID NOS: 1728-1735

C19orf70
Chromosome 19 open reading frame 70
SEQ ID NOS: 1736-1739

C1GALT1
Core 1 synthase, glycoprotein-N-
SEQ ID NOS: 1740-1744

acetylgalactosamine 3-beta-galactosyltransferase 1

C1orf127
Chromosome 1 open reading frame 127
SEQ ID NOS: 1745-1748

C1orf159
Chromosome 1 open reading frame 159
SEQ ID NOS: 1749-1761

C1orf198
Chromosome 1 open reading frame 198
SEQ ID NOS: 1762-1766

C1orf54
Chromosome 1 open reading frame 54
SEQ ID NOS: 1767-1769

C1orf56
Chromosome 1 open reading frame 56
SEQ ID NO: 1770

C1QA
Complement component 1, q subcomponent, A
SEQ ID NOS: 1771-1773

chain

C1QB
Complement component 1, q subcomponent, B
SEQ ID NOS: 1774-1777

chain

C1QC
Complement component 1, q subcomponent, C
SEQ ID NOS: 1778-1780

chain

C1QL1
Complement component 1, q subcomponent-like 1
SEQ ID NO: 1781

C1QL2
Complement component 1, q subcomponent-like 2
SEQ ID NO: 1782

C1QL3
Complement component 1, q subcomponent-like 3
SEQ ID NOS: 1783-1784

C1QL4
Complement component 1, q subcomponent-like 4
SEQ ID NO: 1785

C1QTNF1
C1q and tumor necrosis factor related protein 1
SEQ ID NOS: 1786-1795

FAM132A
Family with sequence similarity 132, member A
SEQ ID NO: 1796

C1QTNF2
C1q and tumor necrosis factor related protein 2
SEQ ID NO: 1797

C1QTNF3
C1q and tumor necrosis factor related protein 3
SEQ ID NOS: 1798-1799

C1QTNF4
C1q and tumor necrosis factor related protein 4
SEQ ID NOS: 1800-1801

C1QTNF5
C1q and tumor necrosis factor related protein 5
SEQ ID NOS: 1802-1804

C1QTNF7
C1q and tumor necrosis factor related protein 7
SEQ ID NOS: 1805-1809

C1QTNF8
C1q and tumor necrosis factor related protein 8
SEQ ID NOS: 1810-1811

C1QTNF9
C1q and tumor necrosis factor related protein 9
SEQ ID NOS: 1812-1813

C1QTNF9B
C1q and tumor necrosis factor related protein 9B
SEQ ID NOS: 1814-1816

C1R
Complement component 1, r subcomponent
SEQ ID NOS: 1817-1825

C1RL
Complement component 1, r subcomponent-like
SEQ ID NOS: 1826-1834

C1S
Complement component 1, s subcomponent
SEQ ID NOS: 1835-1844

C2
Complement component 2
SEQ ID NOS: 1845-1859

C21orf33
Chromosome 21 open reading frame 33
SEQ ID NOS: 1860-1868

C21orf62
Chromosome 21 open reading frame 62
SEQ ID NOS: 1869-1872

C22orf15
Chromosome 22 open reading frame 15
SEQ ID NOS: 1873-1875

C22orf46
Chromosome 22 open reading frame 46
SEQ ID NO: 1876

C2CD2
C2 calcium-dependent domain containing 2
SEQ ID NOS: 1877-1879

C2orf40
Chromosome 2 open reading frame 40
SEQ ID NOS: 1880-1882

C2orf66
Chromosome 2 open reading frame 66
SEQ ID NO: 1883

C2orf69
Chromosome 2 open reading frame 69
SEQ ID NO: 1884

C2orf78
Chromosome 2 open reading frame 78
SEQ ID NO: 1885

C3
Complement component 3
SEQ ID NOS: 1886-1890

C3orf33
Chromosome 3 open reading frame 33
SEQ ID NOS: 1891-1895

C3orf58
Chromosome 3 open reading frame 58
SEQ ID NOS: 1896-1899

C4A
Complement component 4A (Rodgers blood group)
SEQ ID NOS: 1900-1901

C4B
Complement component 4B (Chido blood group)
SEQ ID NOS: 1902-1903

C4BPA
Complement component 4 binding protein, alpha
SEQ ID NOS: 1904-1906

C4BPB
Complement component 4 binding protein, beta
SEQ ID NOS: 1907-1911

C4orf48
Chromosome 4 open reading frame 48
SEQ ID NOS: 1912-1913

C5
Complement component 5
SEQ ID NO: 1914

C5orf46
Chromosome 5 open reading frame 46
SEQ ID NOS: 1915-1916

C6
Complement component 6
SEQ ID NOS: 1917-1920

C6orf120
Chromosome 6 open reading frame 120
SEQ ID NO: 1921

C6orf15
Chromosome 6 open reading frame 15
SEQ ID NO: 1922

C6orf58
Chromosome 6 open reading frame 58
SEQ ID NO: 1923

C7
Complement component 7
SEQ ID NO: 1924

C7orf57
Chromosome 7 open reading frame 57
SEQ ID NOS: 1925-1929

C8A
Complement component 8, alpha polypeptide
SEQ ID NO: 1930

C8B
Complement component 8, beta polypeptide
SEQ ID NOS: 1931-1933

C8G
Complement component 8, gamma polypeptide
SEQ ID NOS: 1934-1935

C9
Complement component 9
SEQ ID NO: 1936

C9orf47
Chromosome 9 open reading frame 47
SEQ ID NOS: 1937-1939

CA10
Carbonic anhydrase X
SEQ ID NOS: 1940-1946

CA11
Carbonic anhydrase XI
SEQ ID NOS: 1947-1948

CA6
Carbonic anhydrase VI
SEQ ID NOS: 1949-1953

CA9
Carbonic anhydrase IX
SEQ ID NOS: 1954-1955

CABLES1
Cdk5 and Abl enzyme substrate 1
SEQ ID NOS: 1956-1961

CABP1
Calcium binding protein 1
SEQ ID NOS: 1962-1965

CACNA2D1
Calcium channel, voltage-dependent, alpha 2/delta
SEQ ID NOS: 1966-1969

subunit 1

CACNA2D4
Calcium channel, voltage-dependent, alpha 2/delta
SEQ ID NOS: 1970-1983

subunit 4

CADM3
Cell adhesion molecule 3
SEQ ID NOS: 1984-1986

CALCA
Calcitonin-related polypeptide alpha
SEQ ID NOS: 1987-1991

CALCB
Calcitonin-related polypeptide beta
SEQ ID NOS: 1992-1994

CALCR
Calcitonin receptor
SEQ ID NOS: 1995-2001

CALCRL
Calcitonin receptor-like
SEQ ID NOS: 2002-2006

FAM26D
Family with sequence similarity 26, member D
SEQ ID NOS: 2007-2011

CALR
Calreticulin
SEQ ID NOS: 2012-2015

CALR3
Calreticulin 3
SEQ ID NOS: 2016-2017

CALU
Calumenin
SEQ ID NOS: 2018-2023

CAMK2D
Calcium/calmodulin-dependent protein kinase II
SEQ ID NOS: 2024-2035

delta

CAMP
Cathelicidin antimicrobial peptide
SEQ ID NO: 2036

CANX
Calnexin
SEQ ID NOS: 2037-2051

CARM1
Coactivator-associated arginine methyltransferase 1
SEQ ID NOS: 2052-2059

CARNS1
Carnosine synthase 1
SEQ ID NOS: 2060-2062

CARTPT
CART prepropeptide
SEQ ID NO: 2063

CASQ1
Calsequestrin 1 (fast-twitch, skeletal muscle)
SEQ ID NOS: 2064-2065

CASQ2
Calsequestrin 2 (cardiac muscle)
SEQ ID NO: 2066

CATSPERG
Catsper channel auxiliary subunit gamma
SEQ ID NOS: 2067-2074

CBLN1
Cerebellin 1 precursor
SEQ ID NOS: 2075-2077

CBLN2
Cerebellin 2 precursor
SEQ ID NOS: 2078-2081

CBLN3
Cerebellin 3 precursor
SEQ ID NOS: 2082-2083

CBLN4
Cerebellin 4 precursor
SEQ ID NO: 2084

CCBE1
Collagen and calcium binding EGF domains 1
SEQ ID NOS: 2085-2087

CCDC112
Coiled-coil domain containing 112
SEQ ID NOS: 2088-2091

CCDC129
Coiled-coil domain containing 129
SEQ ID NOS: 2092-2099

CCDC134
Coiled-coil domain containing 134
SEQ ID NOS: 2100-2101

CCDC149
Coiled-coil domain containing 149
SEQ ID NOS: 2102-2105

CCDC3
Coiled-coil domain containing 3
SEQ ID NOS: 2106-2107

CCDC80
Coiled-coil domain containing 80
SEQ ID NOS: 2108-2111

CCDC85A
Coiled-coil domain containing 85A
SEQ ID NO: 2112

CCDC88B
Coiled-coil domain containing 88B
SEQ ID NOS: 2113-2115

CCER2
Coiled-coil glutamate-rich protein 2
SEQ ID NOS: 2116-2117

CCK
Cholecystokinin
SEQ ID NOS: 2118-2120

CCL1
Chemokine (C-C motif) ligand 1
SEQ ID NO: 2121

CCL11
Chemokine (C-C motif) ligand 11
SEQ ID NO: 2122

CCL13
Chemokine (C-C motif) ligand 13
SEQ ID NOS: 2123-2124

CCL14
Chemokine (C-C motif) ligand 14
SEQ ID NOS: 2125-2128

CCL15
Chemokine (C-C motif) ligand 15
SEQ ID NOS: 2129-2130

CCL16
Chemokine (C-C motif) ligand 16
SEQ ID NOS: 2131-2133

CCL17
Chemokine (C-C motif) ligand 17
SEQ ID NOS: 2134-2135

CCL18
Chemokine (C-C motif) ligand 18 (pulmonary and
SEQ ID NO: 2136

activation-regulated)

CCL19
Chemokine (C-C motif) ligand 19
SEQ ID NOS: 2137-2138

CCL2
Chemokine (C-C motif) ligand 2
SEQ ID NOS: 2139-2140

CCL20
Chemokine (C-C motif) ligand 20
SEQ ID NOS: 2141-2143

CCL21
Chemokine (C-C motif) ligand 21
SEQ ID NOS: 2144-2145

CCL22
Chemokine (C-C motif) ligand 22
SEQ ID NO: 2146

CCL23
Chemokine (C-C motif) ligand 23
SEQ ID NOS: 2147-2149

CCL24
Chemokine (C-C motif) ligand 24
SEQ ID NOS: 2150-2151

CCL25
Chemokine (C-C motif) ligand 25
SEQ ID NOS: 2152-2155

CCL26
Chemokine (C-C motif) ligand 26
SEQ ID NOS: 2156-2157

CCL27
Chemokine (C-C motif) ligand 27
SEQ ID NO: 2158

CCL28
Chemokine (C-C motif) ligand 28
SEQ ID NOS: 2159-2161

CCL3
Chemokine (C-C motif) ligand 3
SEQ ID NO: 2162

CCL3L3
Chemokine (C-C motif) ligand 3-like 3
SEQ ID NO: 2163

CCL4
Chemokine (C-C motif) ligand 4
SEQ ID NOS: 2164-2165

CCL4L2
Chemokine (C-C motif) ligand 4-like 2
SEQ ID NOS: 2166-2175

CCL5
Chemokine (C-C motif) ligand 5
SEQ ID NOS: 2176-2178

CCL7
Chemokine (C-C motif) ligand 7
SEQ ID NOS: 2179-2181

CCL8
Chemokine (C-C motif) ligand 8
SEQ ID NO: 2182

CCNB1IP1
Cyclin B1 interacting protein 1, E3 ubiquitin protein
SEQ ID NOS: 2183-2194

ligase

CCNL1
Cyclin L1
SEQ ID NOS: 2195-2203

CCNL2
Cyclin L2
SEQ ID NOS: 2204-2211

CD14
CD14 molecule
SEQ ID NOS: 2212-2216

CD160
CD160 molecule
SEQ ID NOS: 2217-2221

CD164
CD164 molecule, sialomucin
SEQ ID NOS: 2222-2227

CD177
CD177 molecule
SEQ ID NOS: 2228-2230

CD1E
CD1e molecule
SEQ ID NOS: 2231-2244

CD2
CD2 molecule
SEQ ID NOS: 2245-2246

CD200
CD200 molecule
SEQ ID NOS: 2247-2253

CD200R1
CD200 receptor 1
SEQ ID NOS: 2254-2258

CD22
CD22 molecule
SEQ ID NOS: 2259-2276

CD226
CD226 molecule
SEQ ID NOS: 2277-2284

CD24
CD24 molecule
SEQ ID NOS: 2285-2291

CD276
CD276 molecule
SEQ ID NOS: 2292-2307

CD300A
CD300a molecule
SEQ ID NOS: 2308-2312

CD300LB
CD300 molecule-like family member b
SEQ ID NOS: 2313-2314

CD300LF
CD300 molecule-like family member f
SEQ ID NOS: 2315-2323

CD300LG
CD300 molecule-like family member g
SEQ ID NOS: 2324-2329

CD3D
CD3d molecule, delta (CD3-TCR complex)
SEQ ID NOS: 2330-2333

CD4
CD4 molecule
SEQ ID NOS: 2334-2336

CD40
CD40 molecule, TNF receptor superfamily member 5
SEQ ID NOS: 2337-2340

CD44
CD44 molecule (Indian blood group)
SEQ ID NOS: 2341-2367

CD48
CD48 molecule
SEQ ID NOS: 2368-2370

CD5
CD5 molecule
SEQ ID NOS: 2371-2372

CD55
CD55 molecule, decay accelerating factor for
SEQ ID NOS: 2373-2383

complement (Cromer blood group)

CD59
CD59 molecule, complement regulatory protein
SEQ ID NOS: 2384-2394

CD5L
CD5 molecule-like
SEQ ID NO: 2395

CD6
CD6 molecule
SEQ ID NOS: 2396-2403

CD68
CD68 molecule
SEQ ID NOS: 2404-2407

CD7
CD7 molecule
SEQ ID NOS: 2408-2413

CD79A
CD79a molecule, immunoglobulin-associated alpha
SEQ ID NOS: 2414-2416

CD80
CD80 molecule
SEQ ID NOS: 2417-2419

CD86
CD86 molecule
SEQ ID NOS: 2420-2426

CD8A
CD8a molecule
SEQ ID NOS: 2427-2430

CD8B
CD8b molecule
SEQ ID NOS: 2431-2436

CD99
CD99 molecule
SEQ ID NOS: 2437-2445

CDC23
Cell division cycle 23
SEQ ID NOS: 2446-2450

CDC40
Cell division cycle 40
SEQ ID NOS: 2451-2453

CDC45
Cell division cycle 45
SEQ ID NOS: 2454-2460

CDCP1
CUB domain containing protein 1
SEQ ID NOS: 2461-2462

CDCP2
CUB domain containing protein 2
SEQ ID NOS: 2463-2464

CDH1
Cadherin 1, type 1
SEQ ID NOS: 2465-2472

CDH11
Cadherin 11, type 2, OB-cadherin (osteoblast)
SEQ ID NOS: 2473-2482

CDH13
Cadherin 13
SEQ ID NOS: 2483-2492

CDH17
Cadherin 17, LI cadherin (liver-intestine)
SEQ ID NOS: 2493-2497

CDH18
Cadherin 18, type 2
SEQ ID NOS: 2498-2504

CDH19
Cadherin 19, type 2
SEQ ID NOS: 2505-2509

CDH23
Cadherin-related 23
SEQ ID NOS: 2510-2525

CDH5
Cadherin 5, type 2 (vascular endothelium)
SEQ ID NOS: 2526-2533

CDHR1
Cadherin-related family member 1
SEQ ID NOS: 2534-2539

CDHR4
Cadherin-related family member 4
SEQ ID NOS: 2540-2544

CDHR5
Cadherin-related family member 5
SEQ ID NOS: 2545-2551

CDKN2A
Cyclin-dependent kinase inhibitor 2A
SEQ ID NOS: 2552-2562

CDNF
Cerebral dopamine neurotrophic factor
SEQ ID NOS: 2563-2564

CDON
Cell adhesion associated, oncogene regulated
SEQ ID NOS: 2565-2572

CDSN
Corneodesmosin
SEQ ID NO: 2573

CEACAM16
Carcinoembryonic antigen-related cell adhesion
SEQ ID NOS: 2574-2575

molecule 16

CEACAM18
Carcinoembryonic antigen-related cell adhesion
SEQ ID NO: 2576

molecule 18

CEACAM19
Carcinoembryonic antigen-related cell adhesion
SEQ ID NOS: 2577-2583

molecule 19

CEACAM5
Carcinoembryonic antigen-related cell adhesion
SEQ ID NOS: 2584-2591

molecule 5

CEACAM7
Carcinoembryonic antigen-related cell adhesion
SEQ ID NOS: 2592-2594

molecule 7

CEACAM8
Carcinoembryonic antigen-related cell adhesion
SEQ ID NOS: 2595-2596

molecule 8

CEL
Carboxyl ester lipase
SEQ ID NO: 2597

CELA2A
Chymotrypsin-like elastase family, member 2A
SEQ ID NO: 2598

CELA2B
Chymotrypsin-like elastase family, member 2B
SEQ ID NOS: 2599-2600

CELA3A
Chymotrypsin-like elastase family, member 3A
SEQ ID NOS: 2601-2603

CELA3B
Chymotrypsin-like elastase family, member 3B
SEQ ID NOS: 2604-2606

CEMIP
Cell migration inducing protein, hyaluronan binding
SEQ ID NOS: 2607-2611

CEP89
Centrosomal protein 89 kDa
SEQ ID NOS: 2612-2617

CER1
Cerberus 1, DAN family BMP antagonist
SEQ ID NO: 2618

CERCAM
Cerebral endothelial cell adhesion molecule
SEQ ID NOS: 2619-2626

CERS1
Ceramide synthase 1
SEQ ID NOS: 2627-2631

CES1
Carboxylesterase 1
SEQ ID NOS: 2632-2637

CES3
Carboxylesterase 3
SEQ ID NOS: 2638-2642

CES4A
Carboxylesterase 4A
SEQ ID NOS: 2643-2648

CES5A
Carboxylesterase 5A
SEQ ID NOS: 2649-2656

CETP
Cholesteryl ester transfer protein, plasma
SEQ ID NOS: 2657-2659

CCDC108
Coiled-coil domain containing 108
SEQ ID NOS: 2660-2669

CFB
Complement factor B
SEQ ID NOS: 2670-2674

CFC1
Cripto, FRL-1, cryptic family 1
SEQ ID NOS: 2675-2677

CFC1B
Cripto, FRL-1, cryptic family 1B
SEQ ID NOS: 2678-2680

CFD
Complement factor D (adipsin)
SEQ ID NOS: 2681-2682

CFDP1
Craniofacial development protein 1
SEQ ID NOS: 2683-2686

CFH
Complement factor H
SEQ ID NOS: 2687-2689

CFHR1
Complement factor H-related 1
SEQ ID NOS: 2690-2691

CFHR2
Complement factor H-related 2
SEQ ID NOS: 2692-2693

CFHR3
Complement factor H-related 3
SEQ ID NOS: 2694-2698

CFHR4
Complement factor H-related 4
SEQ ID NOS: 2699-2702

CFHR5
Complement factor H-related 5
SEQ ID NO: 2703

CFI
Complement factor I
SEQ ID NOS: 2704-2708

CFP
Complement factor properdin
SEQ ID NOS: 2709-2712

CGA
Glycoprotein hormones, alpha polypeptide
SEQ ID NOS: 2713-2717

CGB1
Chorionic gonadotropin, beta polypeptide 1
SEQ ID NOS: 2718-2719

CGB2
Chorionic gonadotropin, beta polypeptide 2
SEQ ID NOS: 2720-2721

CGB
Chorionic gonadotropin, beta polypeptide
SEQ ID NO: 2722

CGB5
Chorionic gonadotropin, beta polypeptide 5
SEQ ID NO: 2723

CGB7
Chorionic gonadotropin, beta polypeptide 7
SEQ ID NOS: 2724-2726

CGB8
Chorionic gonadotropin, beta polypeptide 8
SEQ ID NO: 2727

CGREF1
Cell growth regulator with EF-hand domain 1
SEQ ID NOS: 2728-2735

CHAD
Chondroadherin
SEQ ID NOS: 2736-2738

CHADL
Chondroadherin-like
SEQ ID NOS: 2739-2741

CHEK2
Checkpoint kinase 2
SEQ ID NOS: 2742-2763

CHGA
Chromogranin A
SEQ ID NOS: 2764-2766

CHGB
Chromogranin B
SEQ ID NOS: 2767-2768

CHI3L1
Chitinase 3-like 1 (cartilage glycoprotein-39)
SEQ ID NOS: 2769-2770

CHI3L2
Chitinase 3-like 2
SEQ ID NOS: 2771-2784

CHIA
Chitinase, acidic
SEQ ID NOS: 2785-2793

CHID1
Chitinase domain containing 1
SEQ ID NOS: 2794-2812

CHIT1
Chitinase 1 (chitotriosidase)
SEQ ID NOS: 2813-2816

CHL1
Cell adhesion molecule L1-like
SEQ ID NOS: 2817-2825

CHN1
Chimerin 1
SEQ ID NOS: 2826-2836

CHPF
Chondroitin polymerizing factor
SEQ ID NOS: 2837-2839

CHPF2
Chondroitin polymerizing factor 2
SEQ ID NOS: 2840-2843

CHRD
Chordin
SEQ ID NOS: 2844-2849

CHRDL1
Chordin-like 1
SEQ ID NOS: 2850-2854

CHRDL2
Chordin-like 2
SEQ ID NOS: 2855-2863

CHRNA2
Cholinergic receptor, nicotinic, alpha 2 (neuronal)
SEQ ID NOS: 2864-2872

CHRNA5
Cholinergic receptor, nicotinic, alpha 5 (neuronal)
SEQ ID NOS: 2873-2876

CHRNB1
Cholinergic receptor, nicotinic, beta 1 (muscle)
SEQ ID NOS: 2877-2882

CHRND
Cholinergic receptor, nicotinic, delta (muscle)
SEQ ID NOS: 2883-2888

CHST1
Carbohydrate (keratan sulfate Gal-6)
SEQ ID NO: 2889

sulfotransferase 1

CHST10
Carbohydrate sulfotransferase 10
SEQ ID NOS: 2890-2897

CHST11
Carbohydrate (chondroitin 4) sulfotransferase 11
SEQ ID NOS: 2898-2902

CHST13
Carbohydrate (chondroitin 4) sulfotransferase 13
SEQ ID NOS: 2903-2904

CHST4
Carbohydrate (N-acetylglucosamine 6-O)
SEQ ID NOS: 2905-2906

sulfotransferase 4

CHST5
Carbohydrate (N-acetylglucosamine 6-O)
SEQ ID NOS: 2907-2908

sulfotransferase 5

CHST6
Carbohydrate (N-acetylglucosamine 6-O)
SEQ ID NOS: 2909-2910

sulfotransferase 6

CHST7
Carbohydrate (N-acetylglucosamine 6-O)
SEQ ID NO: 2911

sulfotransferase 7

CHST8
Carbohydrate (N-acetylgalactosamine 4-0)
SEQ ID NOS: 2912-2915

sulfotransferase 8

CHSY1
Chondroitin sulfate synthase 1
SEQ ID NOS: 2916-2917

CHSY3
Chondroitin sulfate synthase 3
SEQ ID NO: 2918

CHTF8
Chromosome transmission fidelity factor 8
SEQ ID NOS: 2919-2929

CILP
Cartilage intermediate layer protein, nucleotide
SEQ ID NO: 2930

pyrophosphohydrolase

CILP2
Cartilage intermediate layer protein 2
SEQ ID NOS: 2931-2932

CKLF
Chemokine-like factor
SEQ ID NOS: 2933-2938

CKMT1A
Creatine kinase, mitochondrial 1A
SEQ ID NOS: 2939-2944

CKMT1B
Creatine kinase, mitochondrial 1B
SEQ ID NOS: 2945-2954

CLCA1
Chloride channel accessory 1
SEQ ID NOS: 2955-2956

CLCF1
Cardiotrophin-like cytokine factor 1
SEQ ID NOS: 2957-2958

CLDN15
Claudin 15
SEQ ID NOS: 2959-2964

CLDN7
Claudin 7
SEQ ID NOS: 2965-2971

CLDND1
Claudin domain containing 1
SEQ ID NOS: 2972-2997

CLEC11A
C-type lectin domain family 11, member A
SEQ ID NOS: 2998-3000

CLEC16A
C-type lectin domain family 16, member A
SEQ ID NOS: 3001-3006

CLEC18A
C-type lectin domain family 18, member A
SEQ ID NOS: 3007-3012

CLEC18B
C-type lectin domain family 18, member B
SEQ ID NOS: 3013-3016

CLEC18C
C-type lectin domain family 18, member C
SEQ ID NOS: 3017-3023

CLEC19A
C-type lectin domain family 19, member A
SEQ ID NOS: 3024-3027

CLEC2B
C-type lectin domain family 2, member B
SEQ ID NOS: 3028-3029

CLEC3A
C-type lectin domain family 3, member A
SEQ ID NOS: 3030-3031

CLEC3B
C-type lectin domain family 3, member B
SEQ ID NOS: 3032-3033

CLGN
Calmegin
SEQ ID NOS: 3034-3036

CLN5
Ceroid-lipofuscinosis, neuronal 5
SEQ ID NOS: 3037-3048

CLPS
Colipase, pancreatic
SEQ ID NOS: 3049-3051

CLPSL1
Colipase-like 1
SEQ ID NOS: 3052-3053

CLPSL2
Colipase-like 2
SEQ ID NOS: 3054-3055

CLPX
Caseinolytic mitochondrial matrix peptidase
SEQ ID NOS: 3056-3058

chaperone subunit

CLSTN3
Calsyntenin 3
SEQ ID NOS: 3059-3065

CLU
Clusterin
SEQ ID NOS: 3066-3079

CLUL1
Clusterin-like 1 (retinal)
SEQ ID NOS: 3080-3087

CMA1
Chymase 1, mast cell
SEQ ID NOS: 3088-3089

CMPK1
Cytidine monophosphate (UMP-CMP) kinase 1,
SEQ ID NOS: 3090-3093

cytosolic

CNBD1
Cyclic nucleotide binding domain containing 1
SEQ ID NOS: 3094-3097

CNDP1
Carnosine dipeptidase 1 (metallopeptidase M20
SEQ ID NOS: 3098-3100

family)

RQCD1
RCD1 required for cell differentiation1 homolog (S.
SEQ ID NOS: 3101-3107

pombe)

CNPY2
Canopy FGF signaling regulator 2
SEQ ID NOS: 3108-3112

CNPY3
Canopy FGF signaling regulator 3
SEQ ID NOS: 3113-3114

CNPY4
Canopy FGF signaling regulator 4
SEQ ID NOS: 3115-3117

CNTFR
Ciliary neurotrophic factor receptor
SEQ ID NOS: 3118-3121

CNTN1
Contactin 1
SEQ ID NOS: 3122-3131

CNTN2
Contactin 2 (axonal)
SEQ ID NOS: 3132-3143

CNTN3
Contactin 3 (plasmacytoma associated)
SEQ ID NO: 3144

CNTN4
Contactin 4
SEQ ID NOS: 3145-3153

CNTN5
Contactin 5
SEQ ID NOS: 3154-3159

CNTNAP2
Contactin associated protein-like 2
SEQ ID NOS: 3160-3163

CNTNAP3
Contactin associated protein-like 3
SEQ ID NOS: 3164-3168

CNTNAP3B
Contactin associated protein-like 3B
SEQ ID NOS: 3169-3177

COASY
CoA synthase
SEQ ID NOS: 3178-3187

COCH
Cochlin
SEQ ID NOS: 3188-3199

COG3
Component of oligomeric golgi complex 3
SEQ ID NOS: 3200-3203

COL10A1
Collagen, type X, alpha 1
SEQ ID NOS: 3204-3207

COL11A1
Collagen, type XI, alpha 1
SEQ ID NOS: 3208-3218

COL11A2
Collagen, type XI, alpha 2
SEQ ID NOS: 3219-3223

COL12A1
Collagen, type XII, alpha 1
SEQ ID NOS: 3224-3231

COL14A1
Collagen, type XIV, alpha 1
SEQ ID NOS: 3232-3239

COL15A1
Collagen, type XV, alpha 1
SEQ ID NOS: 3240-3241

COL16A1
Collagen, type XVI, alpha 1
SEQ ID NOS: 3242-3246

COL18A1
Collagen, type XVIII, alpha 1
SEQ ID NOS: 3247-3251

COL19A1
Collagen, type XIX, alpha 1
SEQ ID NOS: 3252-3254

COL1A1
Collagen, type I, alpha 1
SEQ ID NOS: 3255-3256

COL1A2
Collagen, type I, alpha 2
SEQ ID NOS: 3257-3258

COL20A1
Collagen, type XX, alpha 1
SEQ ID NOS: 3259-3262

COL21A1
Collagen, type XXI, alpha 1
SEQ ID NOS: 3263-3268

COL22A1
Collagen, type XXII, alpha 1
SEQ ID NOS: 3269-3271

COL24A1
Collagen, type XXIV, alpha 1
SEQ ID NOS: 3272-3275

COL26A1
Collagen, type XXVI, alpha 1
SEQ ID NOS: 3276-3277

COL27A1
Collagen, type XXVII, alpha 1
SEQ ID NOS: 3278-3280

COL28A1
Collagen, type XXVIII, alpha 1
SEQ ID NOS: 3281-3285

COL2A1
Collagen, type II, alpha 1
SEQ ID NOS: 3286-3287

COL3A1
Collagen, type III, alpha 1
SEQ ID NOS: 3288-3290

COL4A1
Collagen, type IV, alpha 1
SEQ ID NOS: 3291-3293

COL4A2
Collagen, type IV, alpha 2
SEQ ID NOS: 3294-3296

COL4A3
Collagen, type IV, alpha 3 (Goodpasture antigen)
SEQ ID NOS: 3297-3300

COL4A4
Collagen, type IV, alpha 4
SEQ ID NOS: 3301-3302

COL4A5
Collagen, type IV, alpha 5
SEQ ID NOS: 3303-3309

COL4A6
Collagen, type IV, alpha 6
SEQ ID NOS: 3310-3315

COL5A1
Collagen, type V, alpha 1
SEQ ID NOS: 3316-3318

COL5A2
Collagen, type V, alpha 2
SEQ ID NOS: 3319-3320

COL5A3
Collagen, type V, alpha 3
SEQ ID NO: 3321

COL6A1
Collagen, type VI, alpha 1
SEQ ID NOS: 3322-3323

COL6A2
Collagen, type VI, alpha 2
SEQ ID NOS: 3324-3329

COL6A3
Collagen, type VI, alpha 3
SEQ ID NOS: 3330-3338

COL6A5
Collagen, type VI, alpha 5
SEQ ID NOS: 3339-3343

COL6A6
Collagen, type VI, alpha 6
SEQ ID NOS: 3344-3346

COL7A1
Collagen, type VII, alpha 1
SEQ ID NOS: 3347-3348

COL8A1
Collagen, type VIII, alpha 1
SEQ ID NOS: 3349-3352

COL8A2
Collagen, type VIII, alpha 2
SEQ ID NOS: 3353-3355

COL9A1
Collagen, type IX, alpha 1
SEQ ID NOS: 3356-3359

COL9A2
Collagen, type IX, alpha 2
SEQ ID NOS: 3360-3363

COL9A3
Collagen, type IX, alpha 3
SEQ ID NOS: 3364-3365

COLEC10
Collectin sub-family member 10 (C-type lectin)
SEQ ID NO: 3366

COLEC11
Collectin sub-family member 11
SEQ ID NOS: 3367-3376

COLGALT1
Collagen beta(1-O)galactosyltransferase 1
SEQ ID NOS: 3377-3379

COLGALT2
Collagen beta(1-O)galactosyltransferase 2
SEQ ID NOS: 3380-3382

COLQ
Collagen-like tail subunit (single strand of
SEQ ID NOS: 3383-3387

homotrimer) of asymmetric acetylcholinesterase

COMP
Cartilage oligomeric matrix protein
SEQ ID NOS: 3388-3390

COPS6
COP9 signalosome subunit 6
SEQ ID NOS: 3391-3394

COQ6
Coenzyme Q6 monooxygenase
SEQ ID NOS: 3395-3402

CORT
Cortistatin
SEQ ID NO: 3403

CP
Ceruloplasmin (ferroxidase)
SEQ ID NOS: 3404-3408

CPA1
Carboxypeptidase A1 (pancreatic)
SEQ ID NOS: 3409-3413

CPA2
Carboxypeptidase A2 (pancreatic)
SEQ ID NOS: 3414-3415

CPA3
Carboxypeptidase A3 (mast cell)
SEQ ID NO: 3416

CPA4
Carboxypeptidase A4
SEQ ID NOS: 3417-3422

CPA6
Carboxypeptidase A6
SEQ ID NOS: 3423-3425

CPAMD8
C3 and PZP-like, alpha-2-macroglobulin domain
SEQ ID NOS: 3426-3431

containing 8

CPB1
Carboxypeptidase B1 (tissue)
SEQ ID NOS: 3432-3436

CPB2
Carboxypeptidase B2 (plasma)
SEQ ID NOS: 3437-3439

CPE
Carboxypeptidase E
SEQ ID NOS: 3440-3444

CPM
Carboxypeptidase M
SEQ ID NOS: 3445-3454

CPN1
Carboxypeptidase N, polypeptide 1
SEQ ID NOS: 3455-3456

CPN2
Carboxypeptidase N, polypeptide 2
SEQ ID NOS: 3457-3458

CPO
Carboxypeptidase O
SEQ ID NO: 3459

CPQ
Carboxypeptidase Q
SEQ ID NOS: 3460-3465

CPVL
Carboxypeptidase, vitellogenic-like
SEQ ID NOS: 3466-3476

CPXM1
Carboxypeptidase X (M14 family), member 1
SEQ ID NO: 3477

CPXM2
Carboxypeptidase X (M14 family), member 2
SEQ ID NOS: 3478-3479

CPZ
Carboxypeptidase Z
SEQ ID NOS: 3480-3483

CR1L
Complement component (3b/4b) receptor 1-like
SEQ ID NOS: 3484-3485

CRB2
Crumbs family member 2
SEQ ID NOS: 3486-3488

CREG1
Cellular repressor of E1A-stimulated genes 1
SEQ ID NO: 3489

CREG2
Cellular repressor of E1A-stimulated genes 2
SEQ ID NO: 3490

CRELD1
Cysteine-rich with EGF-like domains 1
SEQ ID NOS: 3491-3496

CRELD2
Cysteine-rich with EGF-like domains 2
SEQ ID NOS: 3497-3501

CRH
Corticotropin releasing hormone
SEQ ID NO: 3502

CRHBP
Corticotropin releasing hormone binding protein
SEQ ID NOS: 3503-3504

CRHR1
Corticotropin releasing hormone receptor 1
SEQ ID NOS: 3505-3516

CRHR2
Corticotropin releasing hormone receptor 2
SEQ ID NOS: 3517-3523

CRISP1
Cysteine-rich secretory protein 1
SEQ ID NOS: 3524-3527

CRISP2
Cysteine-rich secretory protein 2
SEQ ID NOS: 3528-3530

CRISP3
Cysteine-rich secretory protein 3
SEQ ID NOS: 3531-3534

CRISPLD2
Cysteine-rich secretory protein LCCL domain
SEQ ID NOS: 3535-3542

containing 2

CRLF1
Cytokine receptor-like factor 1
SEQ ID NOS: 3543-3544

CRP
C-reactive protein, pentraxin-related
SEQ ID NOS: 3545-3549

CRTAC1
Cartilage acidic protein 1
SEQ ID NOS: 3550-3554

CRTAP
Cartilage associated protein
SEQ ID NOS: 3555-3556

CRY2
Cryptochrome circadian clock 2
SEQ ID NOS: 3557-3560

CSAD
Cysteine sulfinic acid decarboxylase
SEQ ID NOS: 3561-3573

CSF1
Colony stimulating factor 1 (macrophage)
SEQ ID NOS: 3574-3581

CSF1R
Colony stimulating factor 1 receptor
SEQ ID NOS: 3582-3586

CSF2
Colony stimulating factor 2 (granulocyte-
SEQ ID NO: 3587

macrophage)

CSF2RA
Colony stimulating factor 2 receptor, alpha, low-
SEQ ID NOS: 3588-3599

affinity (granulocyte-macrophage)

CSF3
Colony stimulating factor 3 (granulocyte)
SEQ ID NOS: 3600-3606

CSGALNACT
Chondroitin sulfate N-
SEQ ID NOS: 3607-3615

1
acetylgalactosaminyltransferase 1

CSH1
Chorionic somatomammotropin hormone 1
SEQ ID NOS: 3616-3619

(placental lactogen)

CSH2
Chorionic somatomammotropin hormone 2
SEQ ID NOS: 3620-3624

CSHL1
Chorionic somatomammotropin hormone-like 1
SEQ ID NOS: 3625-3631

CSN1S1
Casein alpha s1
SEQ ID NOS: 3632-3637

CSN2
Casein beta
SEQ ID NO: 3638

CSN3
Casein kappa
SEQ ID NO: 3639

CST1
Cystatin SN
SEQ ID NOS: 3640-3641

CST11
Cystatin 11
SEQ ID NOS: 3642-3643

CST2
Cystatin SA
SEQ ID NO: 3644

CST3
Cystatin C
SEQ ID NOS: 3645-3647

CST4
Cystatin S
SEQ ID NO: 3648

CST5
Cystatin D
SEQ ID NO: 3649

CST6
Cystatin E/M
SEQ ID NO: 3650

CST7
Cystatin F (leukocystatin)
SEQ ID NO: 3651

CST8
Cystatin 8 (cystatin-related epididymal specific)
SEQ ID NOS: 3652-3653

CST9
Cystatin 9 (testatin)
SEQ ID NO: 3654

CST9L
Cystatin 9-like
SEQ ID NO: 3655

CSTL1
Cystatin-like 1
SEQ ID NOS: 3656-3658

CT55
Cancer/testis antigen 55
SEQ ID NOS: 3659-3660

CTBS
Chitobiase, di-N-acetyl-
SEQ ID NOS: 3661-3663

CTGF
Connective tissue growth factor
SEQ ID NO: 3664

CTHRC1
Collagen triple helix repeat containing 1
SEQ ID NOS: 3665-3668

CTLA4
Cytotoxic T-lymphocyte-associated protein 4
SEQ ID NOS: 3669-3672

CTNS
Cystinosin, lysosomal cystine transporter
SEQ ID NOS: 3673-3680

CTRB1
Chymotrypsinogen B1
SEQ ID NOS: 3681-3683

CTRB2
Chymotrypsinogen B2
SEQ ID NOS: 3684-3687

CTRC
Chymotrypsin C (caldecrin)
SEQ ID NOS: 3688-3689

CTRL
Chymotrypsin-like
SEQ ID NOS: 3690-3692

CTSA
Cathepsin A
SEQ ID NOS: 3693-3701

CTSB
Cathepsin B
SEQ ID NOS: 3702-3726

CTSC
Cathepsin C
SEQ ID NOS: 3727-3731

CTSD
Cathepsin D
SEQ ID NOS: 3732-3742

CTSE
Cathepsin E
SEQ ID NOS: 3743-3744

CTSF
Cathepsin F
SEQ ID NOS: 3745-3748

CTSG
Cathepsin G
SEQ ID NO: 3749

CTSH
Cathepsin H
SEQ ID NOS: 3750-3755

CTSK
Cathepsin K
SEQ ID NOS: 3756-3757

CTSL
Cathepsin L
SEQ ID NOS: 3758-3760

CTSO
Cathepsin O
SEQ ID NO: 3761

CTSS
Cathepsin S
SEQ ID NOS: 3762-3766

CTSV
Cathepsin V
SEQ ID NOS: 3767-3768

CTSW
Cathepsin W
SEQ ID NOS: 3769-3771

CTSZ
Cathepsin Z
SEQ ID NO: 3772

CUBN
Cubilin (intrinsic factor-cobalamin receptor)
SEQ ID NOS: 3773-3776

CUTA
CutA divalent cation tolerance homolog (E. coli)
SEQ ID NOS: 3777-3786

CX3CL1
Chemokine (C-X3-C motif) ligand 1
SEQ ID NOS: 3787-3790

CXADR
Coxsackie virus and adenovirus receptor
SEQ ID NOS: 3791-3795

CXCL1
Chemokine (C-X-C motif) ligand 1 (melanoma growth
SEQ ID NO: 3796

stimulating activity, alpha)

CXCL10
Chemokine (C-X-C motif) ligand 10
SEQ ID NO: 3797

CXCL11
Chemokine (C-X-C motif) ligand 11
SEQ ID NOS: 3798-3799

CXCL12
Chemokine (C-X-C motif) ligand 12
SEQ ID NOS: 3800-3805

CXCL13
Chemokine (C-X-C motif) ligand 13
SEQ ID NO: 3806

CXCL14
Chemokine (C-X-C motif) ligand 14
SEQ ID NOS: 3807-3808

CXCL17
Chemokine (C-X-C motif) ligand 17
SEQ ID NOS: 3809-3810

CXCL2
Chemokine (C-X-C motif) ligand 2
SEQ ID NO: 3811

CXCL3
Chemokine (C-X-C motif) ligand 3
SEQ ID NO: 3812

CXCL5
Chemokine (C-X-C motif) ligand 5
SEQ ID NO: 3813

CXCL6
Chemokine (C-X-C motif) ligand 6
SEQ ID NOS: 3814-3815

CXCL8
Chemokine (C-X-C motif) ligand 8
SEQ ID NOS: 3816-3817

CXCL9
Chemokine (C-X-C motif) ligand 9
SEQ ID NO: 3818

CXorf36
Chromosome X open reading frame 36
SEQ ID NOS: 3819-3820

CYB5D2
Cytochrome b5 domain containing 2
SEQ ID NOS: 3821-3824

CYHR1
Cysteine/histidine-rich 1
SEQ ID NOS: 3825-3832

CYP17A1
Cytochrome P450, family 17, subfamily A,
SEQ ID NOS: 3833-3837

polypeptide 1

CYP20A1
Cytochrome P450, family 20, subfamily A,
SEQ ID NOS: 3838-3844

polypeptide 1

CYP21A2
Cytochrome P450, family 21, subfamily A,
SEQ ID NOS: 3845-3852

polypeptide 2

CYP26B1
Cytochrome P450, family 26, subfamily B,
SEQ ID NOS: 3853-3857

polypeptide 1

CYP2A6
Cytochrome P450, family 2, subfamily A,
SEQ ID NOS: 3858-3859

polypeptide 6

CYP2A7
Cytochrome P450, family 2, subfamily A,
SEQ ID NOS: 3860-3862

polypeptide 7

CYP2B6
Cytochrome P450, family 2, subfamily B,
SEQ ID NOS: 3863-3866

polypeptide 6

CYP2C18
Cytochrome P450, family 2, subfamily C,
SEQ ID NOS: 3867-3868

polypeptide 18

CYP2C19
Cytochrome P450, family 2, subfamily C,
SEQ ID NOS: 3869-3870

polypeptide 19

CYP2C8
Cytochrome P450, family 2, subfamily C,
SEQ ID NOS: 3871-3878

polypeptide 8

CYP2C9
Cytochrome P450, family 2, subfamily C,
SEQ ID NOS: 3879-3881

polypeptide 9

CYP2E1
Cytochrome P450, family 2, subfamily E,
SEQ ID NOS: 3882-3887

polypeptide 1

CYP2F1
Cytochrome P450, family 2, subfamily F,
SEQ ID NOS: 3888-3891

polypeptide 1

CYP2J2
Cytochrome P450, family 2, subfamily J,
SEQ ID NO: 3892

polypeptide 2

CYP2R1
Cytochrome P450, family 2, subfamily R,
SEQ ID NOS: 3893-3898

polypeptide 1

CYP2S1
Cytochrome P450, family 2, subfamily S,
SEQ ID NOS: 3899-3904

polypeptide 1

CYP2W1
Cytochrome P450, family 2, subfamily W,
SEQ ID NOS: 3905-3907

polypeptide 1

CYP46A1
Cytochrome P450, family 46, subfamily A,
SEQ ID NOS: 3908-3912

polypeptide 1

CYP4F11
Cytochrome P450, family 4, subfamily F,
SEQ ID NOS: 3913-3917

polypeptide 11

CYP4F2
Cytochrome P450, family 4, subfamily F,
SEQ ID NOS: 3918-3922

polypeptide 2

CYR61
Cysteine-rich, angiogenic inducer, 61
SEQ ID NO: 3923

CYTL1
Cytokine-like 1
SEQ ID NOS: 3924-3926

D2HGDH
D-2-hydroxyglutarate dehydrogenase
SEQ ID NOS: 3927-3935

DAG1
Dystroglycan 1 (dystrophin-associated glycoprotein
SEQ ID NOS: 3936-3950

1)

DAND5
DAN domain family member 5, BMP antagonist
SEQ ID NOS: 3951-3952

DAO
D-amino-acid oxidase
SEQ ID NOS: 3953-3958

DAZAP2
DAZ associated protein 2
SEQ ID NOS: 3959-3967

DBH
Dopamine beta-hydroxylase (dopamine beta-
SEQ ID NOS: 3968-3969

monooxygenase)

DBNL
Drebrin-like
SEQ ID NOS: 3970-3987

DCD
Dermcidin
SEQ ID NOS: 3988-3990

DCN
Decorin
SEQ ID NOS: 3991-4009

DDIAS
DNA damage-induced apoptosis suppressor
SEQ ID NOS: 4010-4019

DDOST
Dolichyl-diphosphooligosaccharide-protein
SEQ ID NOS: 4020-4023

glycosyltransferase subunit (non-catalytic)

DDR1
Discoidin domain receptor tyrosine kinase 1
SEQ ID NOS: 4024-4069

DDR2
Discoidin domain receptor tyrosine kinase 2
SEQ ID NOS: 4070-4075

DDT
D-dopachrome tautomerase
SEQ ID NOS: 4076-4081

DDX17
DEAD (Asp-Glu-Ala-Asp) box helicase 17
SEQ ID NOS: 4082-4086

DDX20
DEAD (Asp-Glu-Ala-Asp) box polypeptide 20
SEQ ID NOS: 4087-4089

DDX25
DEAD (Asp-Glu-Ala-Asp) box helicase 25
SEQ ID NOS: 4090-4096

DDX28
DEAD (Asp-Glu-Ala-Asp) box polypeptide 28
SEQ ID NO: 4097

DEAF1
DEAF1 transcription factor
SEQ ID NOS: 4098-4100

DEF8
Differentially expressed in FDCP 8 homolog (mouse)
SEQ ID NOS: 4101-4120

DEFA1
Defensin, alpha 1
SEQ ID NOS: 4121-4122

DEFA1B
Defensin, alpha 1B
SEQ ID NO: 4123

DEFA3
Defensin, alpha 3, neutrophil-specific
SEQ ID NO: 4124

DEFA4
Defensin, alpha 4, corticostatin
SEQ ID NO: 4125

DEFA5
Defensin, alpha 5, Paneth cell-specific
SEQ ID NO: 4126

DEFA6
Defensin, alpha 6, Paneth cell-specific
SEQ ID NO: 4127

DEFB1
Defensin, beta 1
SEQ ID NO: 4128

DEFB103A
Defensin, beta 103A
SEQ ID NO: 4129

DEFB103B
Defensin, beta 103B
SEQ ID NO: 4130

DEFB104A
Defensin, beta 104A
SEQ ID NO: 4131

DEFB104B
Defensin, beta 104B
SEQ ID NO: 4132

DEFB105A
Defensin, beta 105A
SEQ ID NO: 4133

DEFB105B
Defensin, beta 105B
SEQ ID NO: 4134

DEFB106A
Defensin, beta 106A
SEQ ID NO: 4135

DEFB106B
Defensin, beta 106B
SEQ ID NO: 4136

DEFB107A
Defensin, beta 107A
SEQ ID NO: 4137

DEFB107B
Defensin, beta 107B
SEQ ID NO: 4138

DEFB108B
Defensin, beta 108B
SEQ ID NO: 4139

DEFB110
Defensin, beta 110
SEQ ID NOS: 4140-4141

DEFB113
Defensin, beta 113
SEQ ID NO: 4142

DEFB114
Defensin, beta 114
SEQ ID NO: 4143

DEFB115
Defensin, beta 115
SEQ ID NO: 4144

DEFB116
Defensin, beta 116
SEQ ID NO: 4145

DEFB118
Defensin, beta 118
SEQ ID NO: 4146

DEFB119
Defensin, beta 119
SEQ ID NOS: 4147-4149

DEFB121
Defensin, beta 121
SEQ ID NO: 4150

DEFB123
Defensin, beta 123
SEQ ID NO: 4151

DEFB124
Defensin, beta 124
SEQ ID NO: 4152

DEFB125
Defensin, beta 125
SEQ ID NO: 4153

DEFB126
Defensin, beta 126
SEQ ID NO: 4154

DEFB127
Defensin, beta 127
SEQ ID NO: 4155

DEFB128
Defensin, beta 128
SEQ ID NO: 4156

DEFB129
Defensin, beta 129
SEQ ID NO: 4157

DEFB130
Defensin, beta 130
SEQ ID NO: 4158

RP11-

SEQ ID NO: 4159

1236K1.1

DEFB131
Defensin, beta 131
SEQ ID NO: 4160

CTD-

SEQ ID NO: 4161

2313N18.7

DEFB132
Defensin, beta 132
SEQ ID NO: 4162

DEFB133
Defensin, beta 133
SEQ ID NO: 4163

DEFB134
Defensin, beta 134
SEQ ID NOS: 4164-4165

DEFB135
Defensin, beta 135
SEQ ID NO: 4166

DEFB136
Defensin, beta 136
SEQ ID NO: 4167

DEFB4A
Defensin, beta 4A
SEQ ID NO: 4168

DEFB4B
Defensin, beta 4B
SEQ ID NO: 4169

C10orf10
Chromosome 10 open reading frame 10
SEQ ID NOS: 4170-4171

DGCR2
DiGeorge syndrome critical region gene 2
SEQ ID NOS: 4172-4175

DHH
Desert hedgehog
SEQ ID NO: 4176

DHRS4
Dehydrogenase/reductase (SDR family) member 4
SEQ ID NOS: 4177-4184

DHRS4L2
Dehydrogenase/reductase (SDR family) member 4
SEQ ID NOS: 4185-4194

like 2

DHRS7
Dehydrogenase/reductase (SDR family) member 7
SEQ ID NOS: 4195-4202

DHRS7C
Dehydrogenase/reductase (SDR family) member 7C
SEQ ID NOS: 4203-4205

DHRS9
Dehydrogenase/reductase (SDR family) member 9
SEQ ID NOS: 4206-4213

DHRSX
Dehydrogenase/reductase (SDR family) X-linked
SEQ ID NOS: 4214-4218

DHX29
DEAH (Asp-Glu-Ala-His) box polypeptide 29
SEQ ID NOS: 4219-4221

DHX30
DEAH (Asp-Glu-Ala-His) box helicase 30
SEQ ID NOS: 4222-4229

DHX8
DEAH (Asp-Glu-Ala-His) box polypeptide 8
SEQ ID NOS: 4230-4234

DIO2
Deiodinase, iodothyronine, type II
SEQ ID NOS: 4235-4244

DIXDC1
DIX domain containing 1
SEQ ID NOS: 4245-4248

DKK1
Dickkopf WNT signaling pathway inhibitor 1
SEQ ID NO: 4249

DKK2
Dickkopf WNT signaling pathway inhibitor 2
SEQ ID NOS: 4250-4252

DKK3
Dickkopf WNT signaling pathway inhibitor 3
SEQ ID NOS: 4253-4258

DKK4
Dickkopf WNT signaling pathway inhibitor 4
SEQ ID NO: 4259

DKKL1
Dickkopf-like 1
SEQ ID NOS: 4260-4265

DLG4
Discs, large homolog 4 (Drosophila)
SEQ ID NOS: 4266-4274

DLK1
Delta-like 1 homolog (Drosophila)
SEQ ID NOS: 4275-4278

DLL1
Delta-like 1 (Drosophila)
SEQ ID NOS: 4279-4280

DLL3
Delta-like 3 (Drosophila)
SEQ ID NOS: 4281-4283

DMBT1
Deleted in malignant brain tumors 1
SEQ ID NOS: 4284-4290

DMKN
Dermokine
SEQ ID NOS: 4291-4337

DMP1
Dentin matrix acidic phosphoprotein 1
SEQ ID NOS: 4338-4339

DMRTA2
DMRT-like family A2
SEQ ID NOS: 4340-4341

DNAAF5
Dynein, axonemal, assembly factor 5
SEQ ID NOS: 4342-4345

DNAH14
Dynein, axonemal, heavy chain 14
SEQ ID NOS: 4346-4360

DNAJB11
DnaJ (Hsp40) homolog, subfamily B, member 11
SEQ ID NOS: 4361-4362

DNAJB9
DnaJ (Hsp40) homolog, subfamily B, member 9
SEQ ID NO: 4363

DNAJC25-
DNAJC25-GNG10 readthrough
SEQ ID NO: 4364

GNG10

DNAJC3
DnaJ (Hsp40) homolog, subfamily C, member 3
SEQ ID NOS: 4365-4366

DNASE1
Deoxyribonuclease I
SEQ ID NOS: 4367-4377

DNASE1L1
Deoxyribonuclease I-like 1
SEQ ID NOS: 4378-4388

DNASE1L2
Deoxyribonuclease I-like 2
SEQ ID NOS: 4389-4394

DNASE1L3
Deoxyribonuclease I-like 3
SEQ ID NOS: 4395-4400

DNASE2
Deoxyribonuclease II, lysosomal
SEQ ID NOS: 4401-4402

DNASE2B
Deoxyribonuclease II beta
SEQ ID NOS: 4403-4404

DPEP1
Dipeptidase 1 (renal)
SEQ ID NOS: 4405-4409

DPEP2
Dipeptidase 2
SEQ ID NOS: 4410-4416

DPEP3
Dipeptidase 3
SEQ ID NO: 4417

DPF3
D4, zinc and double PHD fingers, family 3
SEQ ID NOS: 4418-4424

DPP4
Dipeptidyl-peptidase 4
SEQ ID NOS: 4425-4429

DPP7
Dipeptidyl-peptidase 7
SEQ ID NOS: 4430-4435

DPT
Dermatopontin
SEQ ID NO: 4436

DRAXIN
Dorsal inhibitory axon guidance protein
SEQ ID NO: 4437

DSE
Dermatan sulfate epimerase
SEQ ID NOS: 4438-4446

DSG2
Desmoglein 2
SEQ ID NOS: 4447-4448

DSPP
Dentin sialophosphoprotein
SEQ ID NOS: 4449-4450

DST
Dystonin
SEQ ID NOS: 4451-4469

DUOX1
Dual oxidase 1
SEQ ID NOS: 4470-4474

DYNLT3
Dynein, light chain, Tctex-type 3
SEQ ID NOS: 4475-4477

E2F5
E2F transcription factor 5, p130-binding
SEQ ID NOS: 4478-4484

EBAG9
Estrogen receptor binding site associated, antigen, 9
SEQ ID NOS: 4485-4493

EBI3
Epstein-Barr virus induced 3
SEQ ID NO: 4494

ECHDC1
Ethylmalonyl-CoA decarboxylase 1
SEQ ID NOS: 4495-4513

ECM1
Extracellular matrix protein 1
SEQ ID NOS: 4514-4516

ECM2
Extracellular matrix protein 2, female organ and
SEQ ID NOS: 4517-4520

adipocyte specific

ECSIT
ECSIT signalling integrator
SEQ ID NOS: 4521-4532

EDDM3A
Epididymal protein 3A
SEQ ID NO: 4533

EDDM3B
Epididymal protein 3B
SEQ ID NO: 4534

EDEM2
ER degradation enhancer, mannosidase alpha-like 2
SEQ ID NOS: 4535-4536

EDEM3
ER degradation enhancer, mannosidase alpha-like 3
SEQ ID NOS: 4537-4539

EDIL3
EGF-like repeats and discoidin I-like domains 3
SEQ ID NOS: 4540-4541

EDN1
Endothelin 1
SEQ ID NO: 4542

EDN2
Endothelin 2
SEQ ID NO: 4543

EDN3
Endothelin 3
SEQ ID NOS: 4544-4549

EDNRB
Endothelin receptor type B
SEQ ID NOS: 4550-4558

EFEMP1
EGF containing fibulin-like extracellular matrix
SEQ ID NOS: 4559-4569

protein 1

EFEMP2
EGF containing fibulin-like extracellular matrix
SEQ ID NOS: 4570-4581

protein 2

EFNA1
Ephrin-A1
SEQ ID NOS: 4582-4583

EFNA2
Ephrin-A2
SEQ ID NO: 4584

EFNA4
Ephrin-A4
SEQ ID NOS: 4585-4587

EGFL6
EGF-like-domain, multiple 6
SEQ ID NOS: 4588-4589

EGFL7
EGF-like-domain, multiple 7
SEQ ID NOS: 4590-4594

EGFL8
EGF-like-domain, multiple 8
SEQ ID NOS: 4595-4597

EGFLAM
EGF-like, fibronectin type III and laminin G domains
SEQ ID NOS: 4598-4606

EGFR
Epidermal growth factor receptor
SEQ ID NOS: 4607-4614

EHBP1
EH domain binding protein 1
SEQ ID NOS: 4615-4626

EHF
Ets homologous factor
SEQ ID NOS: 4627-4636

EHMT1
Euchromatic histone-lysine N-methyltransferase 1
SEQ ID NOS: 4637-4662

EHMT2
Euchromatic histone-lysine N-methyltransferase 2
SEQ ID NOS: 4663-4667

EIF2AK1
Eukaryotic translation initiation factor 2-alpha
SEQ ID NOS: 4668-4671

kinase 1

ELANE
Elastase, neutrophil expressed
SEQ ID NOS: 4672-4673

ELN
Elastin
SEQ ID NOS: 4674-4696

ELP2
Elongator acetyltransferase complex subunit 2
SEQ ID NOS: 4697-4709

ELSPBP1
Epididymal sperm binding protein 1
SEQ ID NOS: 4710-4715

EMC1
ER membrane protein complex subunit 1
SEQ ID NOS: 4716-4722

EMC10
ER membrane protein complex subunit 10
SEQ ID NOS: 4723-4729

EMC9
ER membrane protein complex subunit 9
SEQ ID NOS: 4730-4733

EMCN
Endomucin
SEQ ID NOS: 4734-4738

EMID1
EMI domain containing 1
SEQ ID NOS: 4739-4745

EMILIN1
Elastin microfibril interfacer 1
SEQ ID NOS: 4746-4747

EMILIN2
Elastin microfibril interfacer 2
SEQ ID NO: 4748

EMILIN3
Elastin microfibril interfacer 3
SEQ ID NO: 4749

ENAM
Enamelin
SEQ ID NO: 4750

ENDOG
Endonuclease G
SEQ ID NO: 4751

ENDOU
Endonuclease, polyU-specific
SEQ ID NOS: 4752-4754

ENHO
Energy homeostasis associated
SEQ ID NO: 4755

ENO4
Enolase family member 4
SEQ ID NOS: 4756-4760

ENPP6
Ectonucleotide pyrophosphatase/
SEQ ID NOS: 4761-4762

phosphodiesterase 6

ENPP7
Ectonucleotide pyrophosphatase/
SEQ ID NOS: 4763-4764

phosphodiesterase 7

ENTPD5
Ectonucleoside triphosphate diphosphohydrolase 5
SEQ ID NOS: 4765-4769

ENTPD8
Ectonucleoside triphosphate diphosphohydrolase 8
SEQ ID NOS: 4770-4773

EOGT
EGF domain-specific O-linked N-acetylglucosamine
SEQ ID NOS: 4774-4781

(GlcNAc) transferase

EPCAM
Epithelial cell adhesion molecule
SEQ ID NOS: 4782-4785

EPDR1
Ependymin related 1
SEQ ID NOS: 4786-4789

EPGN
Epithelial mitogen
SEQ ID NOS: 4790-4798

EPHA10
EPH receptor A10
SEQ ID NOS: 4799-4806

EPHA3
EPH receptor A3
SEQ ID NOS: 4807-4809

EPHA4
EPH receptor A4
SEQ ID NOS: 4810-4819

EPHA7
EPH receptor A7
SEQ ID NOS: 4820-4821

EPHA8
EPH receptor A8
SEQ ID NOS: 4822-4823

EPHB2
EPH receptor B2
SEQ ID NOS: 4824-4828

EPHB4
EPH receptor B4
SEQ ID NOS: 4829-4831

EPHX3
Epoxide hydrolase 3
SEQ ID NOS: 4832-4835

EPO
Erythropoietin
SEQ ID NO: 4836

EPPIN
Epididymal peptidase inhibitor
SEQ ID NOS: 4837-4839

EPPIN-
EPPIN-WFDC6 readthrough
SEQ ID NO: 4840

WFDC6

EPS15
Epidermal growth factor receptor pathway
SEQ ID NOS: 4841-4843

substrate 15

EPS8L1
EPS8-like 1
SEQ ID NOS: 4844-4849

EPX
Eosinophil peroxidase
SEQ ID NO: 4850

EPYC
Epiphycan
SEQ ID NOS: 4851-4852

EQTN
Equatorin, sperm acrosome associated
SEQ ID NOS: 4853-4855

ERAP1
Endoplasmic reticulum aminopeptidase 1
SEQ ID NOS: 4856-4861

ERAP2
Endoplasmic reticulum aminopeptidase 2
SEQ ID NOS: 4862-4869

ERBB3
Erb-b2 receptor tyrosine kinase 3
SEQ ID NOS: 4870-4883

FAM132B
Family with sequence similarity 132, member B
SEQ ID NOS: 4884-4886

ERLIN1
ER lipid raft associated 1
SEQ ID NOS: 4887-4889

ERLIN2
ER lipid raft associated 2
SEQ ID NOS: 4890-4898

ERN1
Endoplasmic reticulum to nucleus signaling 1
SEQ ID NOS: 4899-4900

ERN2
Endoplasmic reticulum to nucleus signaling 2
SEQ ID NOS: 4901-4905

ERO1A
Endoplasmic reticulum oxidoreductase alpha
SEQ ID NOS: 4906-4912

ERO1B
Endoplasmic reticulum oxidoreductase beta
SEQ ID NOS: 4913-4915

ERP27
Endoplasmic reticulum protein 27
SEQ ID NOS: 4916-4917

ERP29
Endoplasmic reticulum protein 29
SEQ ID NOS: 4918-4921

ERP44
Endoplasmic reticulum protein 44
SEQ ID NO: 4922

ERV3-1
Endogenous retrovirus group 3, member 1
SEQ ID NO: 4923

ESM1
Endothelial cell-specific molecule 1
SEQ ID NOS: 4924-4926

ESRP1
Epithelial splicing regulatory protein 1
SEQ ID NOS: 4927-4935

EXOG
Endo/exonuclease (5′-3′), endonuclease G-like
SEQ ID NOS: 4936-4949

EXTL1
Exostosin-like glycosyltransferase 1
SEQ ID NO: 4950

EXTL2
Exostosin-like glycosyltransferase 2
SEQ ID NOS: 4951-4955

F10
Coagulation factor X
SEQ ID NOS: 4956-4959

F11
Coagulation factor XI
SEQ ID NOS: 4960-4964

F12
Coagulation factor XII (Hageman factor)
SEQ ID NO: 4965

F13B
Coagulation factor XIII, B polypeptide
SEQ ID NO: 4966

F2
Coagulation factor II (thrombin)
SEQ ID NOS: 4967-4969

F2R
Coagulation factor II (thrombin) receptor
SEQ ID NOS: 4970-4971

F2RL3
Coagulation factor II (thrombin) receptor-like 3
SEQ ID NOS: 4972-4973

F5
Coagulation factor V (proaccelerin, labile factor)
SEQ ID NOS: 4974-4975

F7
Coagulation factor VII (serum prothrombin
SEQ ID NOS: 4976-4979

conversion accelerator)

F8
Coagulation factor VIII, procoagulant component
SEQ ID NOS: 4980-4985

F9
Coagulation factor IX
SEQ ID NOS: 4986-4987

FABP6
Fatty acid binding protein 6, ileal
SEQ ID NOS: 4988-4990

FAM107B
Family with sequence similarity 107, member B
SEQ ID NOS: 4991-5012

FAM131A
Family with sequence similarity 131, member A
SEQ ID NOS: 5013-5021

FAM171A1
Family with sequence similarity 171, member A1
SEQ ID NOS: 5022-5023

FAM171B
Family with sequence similarity 171, member B
SEQ ID NOS: 5024-5025

FAM172A
Family with sequence similarity 172, member A
SEQ ID NOS: 5026-5030

FAM177A1
Family with sequence similarity 177, member A1
SEQ ID NOS: 5031-5040

FAM180A
Family with sequence similarity 180, member A
SEQ ID NOS: 5041-5043

FAM189A1
Family with sequence similarity 189, member A1
SEQ ID NOS: 5044-5045

FAM198A
Family with sequence similarity 198, member A
SEQ ID NOS: 5046-5048

FAM19A1
Family with sequence similarity 19 (chemokine (C-C
SEQ ID NOS: 5049-5051

motif)-like), member A1

FAM19A2
Family with sequence similarity 19 (chemokine (C-C
SEQ ID NOS: 5052-5059

motif)-like), member A2

FAM19A3
Family with sequence similarity 19 (chemokine (C-C
SEQ ID NOS: 5060-5061

motif)-like), member A3

FAM19A4
Family with sequence similarity 19 (chemokine (C-C
SEQ ID NOS: 5062-5064

motif)-like), member A4

FAM19A5
Family with sequence similarity 19 (chemokine (C-C
SEQ ID NOS: 5065-5068

motif)-like), member A5

FAM20A
Family with sequence similarity 20, member A
SEQ ID NOS: 5069-5072

FAM20C
Family with sequence similarity 20, member C
SEQ ID NO: 5073

FAM213A
Family with sequence similarity 213, member A
SEQ ID NOS: 5074-5079

FAM46B
Family with sequence similarity 46, member B
SEQ ID NO: 5080

FAM57A
Family with sequence similarity 57, member A
SEQ ID NOS: 5081-5086

FAM78A
Family with sequence similarity 78, member A
SEQ ID NOS: 5087-5089

FAM96A
Family with sequence similarity 96, member A
SEQ ID NOS: 5090-5094

FAM9B
Family with sequence similarity 9, member B
SEQ ID NOS: 5095-5098

FAP
Fibroblast activation protein, alpha
SEQ ID NOS: 5099-5105

FAS
Fas cell surface death receptor
SEQ ID NOS: 5106-5115

FAT1
FAT atypical cadherin 1
SEQ ID NOS: 5116-5122

FBLN1
Fibulin 1
SEQ ID NOS: 5123-5135

FBLN2
Fibulin 2
SEQ ID NOS: 5136-5141

FBLN5
Fibulin 5
SEQ ID NOS: 5142-5147

FBLN7
Fibulin 7
SEQ ID NOS: 5148-5153

FBN1
Fibrillin 1
SEQ ID NOS: 5154-5157

FBN2
Fibrillin 2
SEQ ID NOS: 5158-5163

FBN3
Fibrillin 3
SEQ ID NOS: 5164-5168

FBXW7
F-box and WD repeat domain containing 7, E3
SEQ ID NOS: 5169-5179

ubiquitin protein ligase

FCAR
Fc fragment of IgA receptor
SEQ ID NOS: 5180-5189

FCGBP
Fc fragment of IgG binding protein
SEQ ID NOS: 5190-5192

FCGR1B
Fc fragment of IgG, high affinity Ib, receptor (CD64)
SEQ ID NOS: 5193-5198

FCGR3A
Fc fragment of IgG, low affinity IIIa, receptor (CD16a)
SEQ ID NOS: 5199-5205

FCGRT
Fc fragment of IgG, receptor, transporter, alpha
SEQ ID NOS: 5206-5216

FCMR
Fc fragment of IgM receptor
SEQ ID NOS: 5217-5223

FCN1
Ficolin (collagen/fibrinogen domain containing) 1
SEQ ID NOS: 5224-5225

FCN2
Ficolin (collagen/fibrinogen domain containing
SEQ ID NOS: 5226-5227

lectin) 2

FCN3
Ficolin (collagen/fibrinogen domain containing) 3
SEQ ID NOS: 5228-5229

FCRL1
Fc receptor-like 1
SEQ ID NOS: 5230-5232

FCRL3
Fc receptor-like 3
SEQ ID NOS: 5233-5238

FCRL5
Fc receptor-like 5
SEQ ID NOS: 5239-5241

FCRLA
Fc receptor-like A
SEQ ID NOS: 5242-5253

FCRLB
Fc receptor-like B
SEQ ID NOS: 5254-5258

FDCSP
Follicular dendritic cell secreted protein
SEQ ID NO: 5259

FETUB
Fetuin B
SEQ ID NOS: 5260-5266

FGA
Fibrinogen alpha chain
SEQ ID NOS: 5267-5269

FGB
Fibrinogen beta chain
SEQ ID NOS: 5270-5272

FGF10
Fibroblast growth factor 10
SEQ ID NOS: 5273-5274

FGF17
Fibroblast growth factor 17
SEQ ID NOS: 5275-5276

FGF18
Fibroblast growth factor 18
SEQ ID NO: 5277

FGF19
Fibroblast growth factor 19
SEQ ID NO: 5278

FGF21
Fibroblast growth factor 21
SEQ ID NOS: 5279-5280

FGF22
Fibroblast growth factor 22
SEQ ID NOS: 5281-5282

FGF23
Fibroblast growth factor 23
SEQ ID NO: 5283

FGF3
Fibroblast growth factor 3
SEQ ID NO: 5284

FGF4
Fibroblast growth factor 4
SEQ ID NO: 5285

FGF5
Fibroblast growth factor 5
SEQ ID NOS: 5286-5288

FGF7
Fibroblast growth factor 7
SEQ ID NOS: 5289-5293

FGF8
Fibroblast growth factor 8 (androgen-induced)
SEQ ID NOS: 5294-5299

FGFBP1
Fibroblast growth factor binding protein 1
SEQ ID NO: 5300

FGFBP2
Fibroblast growth factor binding protein 2
SEQ ID NO: 5301

FGFBP3
Fibroblast growth factor binding protein 3
SEQ ID NO: 5302

FGFR1
Fibroblast growth factor receptor 1
SEQ ID NOS: 5303-5325

FGFR2
Fibroblast growth factor receptor 2
SEQ ID NOS: 5326-5347

FGFR3
Fibroblast growth factor receptor 3
SEQ ID NOS: 5348-5355

FGFR4
Fibroblast growth factor receptor 4
SEQ ID NOS: 5356-5365

FGFRL1
Fibroblast growth factor receptor-like 1
SEQ ID NOS: 5366-5371

FGG
Fibrinogen gamma chain
SEQ ID NOS: 5372-5377

FGL1
Fibrinogen-like 1
SEQ ID NOS: 5378-5384

FGL2
Fibrinogen-like 2
SEQ ID NOS: 5385-5386

FHL1
Four and a half LIM domains 1
SEQ ID NOS: 5387-5414

FHOD3
Formin homology 2 domain containing 3
SEQ ID NOS: 5415-5421

FIBIN
Fin bud initiation factor homolog (zebrafish)
SEQ ID NO: 5422

FICD
FIC domain containing
SEQ ID NOS: 5423-5426

FJX1
Four jointed box 1
SEQ ID NO: 5427

FKBP10
FK506 binding protein 10, 65 kDa
SEQ ID NOS: 5428-5433

FKBP11
FK506 binding protein 11, 19 kDa
SEQ ID NOS: 5434-5440

FKBP14
FK506 binding protein 14, 22 kDa
SEQ ID NOS: 5441-5443

FKBP2
FK506 binding protein 2, 13 kDa
SEQ ID NOS: 5444-5447

FKBP7
FK506 binding protein 7
SEQ ID NOS: 5448-5453

FKBP9
FK506 binding protein 9, 63 kDa
SEQ ID NOS: 5454-5457

FLT1
Fms-related tyrosine kinase 1
SEQ ID NOS: 5458-5466

FLT4
Fms-related tyrosine kinase 4
SEQ ID NOS: 5467-5471

FMO1
Flavin containing monooxygenase 1
SEQ ID NOS: 5472-5476

FMO2
Flavin containing monooxygenase 2 (non-functional)
SEQ ID NOS: 5477-5479

FMO3
Flavin containing monooxygenase 3
SEQ ID NOS: 5480-5482

FMO5
Flavin containing monooxygenase 5
SEQ ID NOS: 5483-5489

FMOD
Fibromodulin
SEQ ID NO: 5490

FN1
Fibronectin 1
SEQ ID NOS: 5491-5503

FNDC1
Fibronectin type III domain containing 1
SEQ ID NOS: 5504-5505

FNDC7
Fibronectin type III domain containing 7
SEQ ID NOS: 5506-5507

FOCAD
Focadhesin
SEQ ID NOS: 5508-5514

FOLR2
Folate receptor 2 (fetal)
SEQ ID NOS: 5515-5524

FOLR3
Folate receptor 3 (gamma)
SEQ ID NOS: 5525-5529

FOXRED2
FAD-dependent oxidoreductase domain containing 2
SEQ ID NOS: 5530-5533

FP325331.1
Uncharacterized protein UNQ6126/PRO20091
SEQ ID NO: 5534

CH507-

SEQ ID NOS: 5535-5541

9B2.3

FPGS
Folylpolyglutamate synthase
SEQ ID NOS: 5542-5548

FRAS1
Fraser extracellular matrix complex subunit 1
SEQ ID NOS: 5549-5554

FREM1
FRAS1 related extracellular matrix 1
SEQ ID NOS: 5555-5559

FREM3
FRAS1 related extracellular matrix 3
SEQ ID NO: 5560

FRMPD2
FERM and PDZ domain containing 2
SEQ ID NOS: 5561-5564

FRZB
Frizzled-related protein
SEQ ID NO: 5565

FSHB
Follicle stimulating hormone, beta polypeptide
SEQ ID NOS: 5566-5568

FSHR
Follicle stimulating hormone receptor
SEQ ID NOS: 5569-5572

FST
Follistatin
SEQ ID NOS: 5573-5576

FSTL1
Follistatin-like 1
SEQ ID NOS: 5577-5580

FSTL3
Follistatin-like 3 (secreted glycoprotein)
SEQ ID NOS: 5581-5586

FSTL4
Follistatin-like 4
SEQ ID NOS: 5587-5589

FSTL5
Follistatin-like 5
SEQ ID NOS: 5590-5592

FTCDNL1
Formiminotransferase cyclodeaminase N-terminal
SEQ ID NOS: 5593-5596

like

FUCA1
Fucosidase, alpha-L-1, tissue
SEQ ID NO: 5597

FUCA2
Fucosidase, alpha-L-2, plasma
SEQ ID NOS: 5598-5599

FURIN
Furin (paired basic amino acid cleaving enzyme)
SEQ ID NOS: 5600-5606

FUT10
Fucosyltransferase 10 (alpha (1,3)
SEQ ID NOS: 5607-5609

fucosyltransferase)

FUT11
Fucosyltransferase 11 (alpha (1,3)
SEQ ID NOS: 5610-5611

fucosyltransferase)

FXN
Frataxin
SEQ ID NOS: 5612-5619

FXR1
Fragile X mental retardation, autosomal homolog 1
SEQ ID NOS: 5620-5632

FXYD3
FXYD domain containing ion transport regulator 3
SEQ ID NOS: 5633-5645

GABBR1
Gamma-aminobutyric acid (GABA) B receptor, 1
SEQ ID NOS: 5646-5657

GABRA1
Gamma-aminobutyric acid (GABA) A receptor,
SEQ ID NOS: 5658-5673

alpha 1

GABRA2
Gamma-aminobutyric acid (GABA) A receptor,
SEQ ID NOS: 5674-5688

alpha 2

GABRA5
Gamma-aminobutyric acid (GABA) A receptor,
SEQ ID NOS: 5689-5697

alpha 5

GABRG3
Gamma-aminobutyric acid (GABA) A receptor,
SEQ ID NOS: 5698-5703

gamma 3

GABRP
Gamma-aminobutyric acid (GABA) A receptor, pi
SEQ ID NOS: 5704-5712

GAL
Galanin/GMAP prepropeptide
SEQ ID NO: 5713

GAL3ST1
Galactose-3-O-sulfotransferase 1
SEQ ID NOS: 5714-5735

GAL3ST2
Galactose-3-O-sulfotransferase 2
SEQ ID NO: 5736

GAL3ST3
Galactose-3-O-sulfotransferase 3
SEQ ID NOS: 5737-5738

GALC
Galactosylceramidase
SEQ ID NOS: 5739-5748

GALNS
Galactosamine (N-acetyl)-6-sulfatase
SEQ ID NOS: 5749-5754

GALNT10
Polypeptide N-acetylgalactosaminyltransferase 10
SEQ ID NOS: 5755-5758

GALNT12
Polypeptide N-acetylgalactosaminyltransferase 12
SEQ ID NOS: 5759-5760

GALNT15
Polypeptide N-acetylgalactosaminyltransferase 15
SEQ ID NOS: 5761-5764

GALNT2
Polypeptide N-acetylgalactosaminyltransferase 2
SEQ ID NO: 5765

GALNT6
Polypeptide N-acetylgalactosaminyltransferase 6
SEQ ID NOS: 5766-5777

GALNT8
Polypeptide N-acetylgalactosaminyltransferase 8
SEQ ID NOS: 5778-5781

GALNTL6
Polypeptide N-acetylgalactosaminyltransferase-
SEQ ID NOS: 5782-5785

like 6

GALP
Galanin-like peptide
SEQ ID NOS: 5786-5788

GANAB
Glucosidase, alpha; neutral AB
SEQ ID NOS: 5789-5797

GARS
Glycyl-tRNA synthetase
SEQ ID NOS: 5798-5801

GAS1
Growth arrest-specific 1
SEQ ID NO: 5802

GAS6
Growth arrest-specific 6
SEQ ID NO: 5803

GAST
Gastrin
SEQ ID NO: 5804

PDDC1
Parkinson disease 7 domain containing 1
SEQ ID NOS: 5805-5813

GBA
Glucosidase, beta, acid
SEQ ID NOS: 5814-5817

GBGT1
Globoside alpha-1,3-N-
SEQ ID NOS: 5818-5826

acetylgalactosaminyltransferase 1

GC
Group-specific component (vitamin D binding
SEQ ID NOS: 5827-5831

protein)

GCG
Glucagon
SEQ ID NOS: 5832-5833

GCGR
Glucagon receptor
SEQ ID NOS: 5834-5836

GCNT7
Glucosaminyl (N-acetyl) transferase family
SEQ ID NOS: 5837-5838

member 7

GCSH
Glycine cleavage system protein H (aminomethyl
SEQ ID NOS: 5839-5847

carrier)

GDF1
Growth differentiation factor 1
SEQ ID NO: 5848

GDF10
Growth differentiation factor 10
SEQ ID NO: 5849

GDF11
Growth differentiation factor 11
SEQ ID NOS: 5850-5851

GDF15
Growth differentiation factor 15
SEQ ID NOS: 5852-5854

GDF2
Growth differentiation factor 2
SEQ ID NO: 5855

GDF3
Growth differentiation factor 3
SEQ ID NO: 5856

GDF5
Growth differentiation factor 5
SEQ ID NOS: 5857-5858

GDF6
Growth differentiation factor 6
SEQ ID NOS: 5859-5861

GDF7
Growth differentiation factor 7
SEQ ID NO: 5862

GDF9
Growth differentiation factor 9
SEQ ID NOS: 5863-5867

GDNF
Glial cell derived neurotrophic factor
SEQ ID NOS: 5868-5875

GFOD2
Glucose-fructose oxidoreductase domain
SEQ ID NOS: 5876-5881

containing 2

GFPT2
Glutamine-fructose-6-phosphate transaminase 2
SEQ ID NOS: 5882-5884

GFRA2
GDNF family receptor alpha 2
SEQ ID NOS: 5885-5891

GFRA4
GDNF family receptor alpha 4
SEQ ID NOS: 5892-5894

GGA2
Golgi-associated, gamma adaptin ear containing,
SEQ ID NOS: 5895-5903

ARF binding protein 2

GGH
Gamma-glutamyl hydrolase (conjugase,
SEQ ID NO: 5904

folylpolygammaglutamyl hydrolase)

GGT1
Gamma-glutamyltransferase 1
SEQ ID NOS: 5905-5927

GGT5
Gamma-glutamyltransferase 5
SEQ ID NOS: 5928-5932

GH1
Growth hormone 1
SEQ ID NOS: 5933-5937

GH2
Growth hormone 2
SEQ ID NOS: 5938-5942

GHDC
GH3 domain containing
SEQ ID NOS: 5943-5950

GHRH
Growth hormone releasing hormone
SEQ ID NOS: 5951-5953

GHRHR
Growth hormone releasing hormone receptor
SEQ ID NOS: 5954-5959

GHRL
Ghrelin/obestatin prepropeptide
SEQ ID NOS: 5960-5970

GIF
Gastric intrinsic factor (vitamin B synthesis)
SEQ ID NOS: 5971-5972

GIP
Gastric inhibitory polypeptide
SEQ ID NO: 5973

GKN1
Gastrokine 1
SEQ ID NO: 5974

GKN2
Gastrokine 2
SEQ ID NOS: 5975-5976

GLA
Galactosidase, alpha
SEQ ID NOS: 5977-5978

GLB1
Galactosidase, beta 1
SEQ ID NOS: 5979-5987

GLB1L
Galactosidase, beta 1-like
SEQ ID NOS: 5988-5995

GLB1L2
Galactosidase, beta 1-like 2
SEQ ID NOS: 5996-5997

GLCE
Glucuronic acid epimerase
SEQ ID NOS: 5998-5999

GLG1
Golgi glycoprotein 1
SEQ ID NOS: 6000-6007

GLIPR1
GLI pathogenesis-related 1
SEQ ID NOS: 6008-6011

GLIPR1L1
GLI pathogenesis-related 1 like 1
SEQ ID NOS: 6012-6015

GLIS3
GLIS family zinc finger 3
SEQ ID NOS: 6016-6024

GLMP
Glycosylated lysosomal membrane protein
SEQ ID NOS: 6025-6033

GLRB
Glycine receptor, beta
SEQ ID NOS: 6034-6039

GLS
Glutaminase
SEQ ID NOS: 6040-6047

GLT6D1
Glycosyltransferase 6 domain containing 1
SEQ ID NOS: 6048-6049

GLTPD2
Glycolipid transfer protein domain containing 2
SEQ ID NO: 6050

GLUD1
Glutamate dehydrogenase 1
SEQ ID NO: 6051

GM2A
GM2 ganglioside activator
SEQ ID NOS: 6052-6054

GML
Glycosylphosphatidylinositol anchored molecule like
SEQ ID NOS: 6055-6056

GNAS
GNAS complex locus
SEQ ID NOS: 6057-6078

GNLY
Granulysin
SEQ ID NOS: 6079-6082

GNPTG
N-acetylglucosamine-1-phosphate transferase,
SEQ ID NOS: 6083-6087

gamma subunit

GNRH1
Gonadotropin-releasing hormone 1 (luteinizing-
SEQ ID NOS: 6088-6089

releasing hormone)

GNRH2
Gonadotropin-releasing hormone 2
SEQ ID NOS: 6090-6093

GNS
Glucosamine (N-acetyl)-6-sulfatase
SEQ ID NOS: 6094-6099

GOLM1
Golgi membrane protein 1
SEQ ID NOS: 6100-6104

GORAB
Golgin, RAB6-interacting
SEQ ID NOS: 6105-6107

GOT2
Glutamic-oxaloacetic transaminase 2, mitochondrial
SEQ ID NOS: 6108-6110

GP2
Glycoprotein 2 (zymogen granule membrane)
SEQ ID NOS: 6111-6119

GP6
Glycoprotein VI (platelet)
SEQ ID NOS: 6120-6123

GPC2
Glypican 2
SEQ ID NOS: 6124-6125

GPC5
Glypican 5
SEQ ID NOS: 6126-6128

GPC6
Glypican 6
SEQ ID NOS: 6129-6130

GPD2
Glycerol-3-phosphate dehydrogenase 2
SEQ ID NOS: 6131-6139

(mitochondrial)

GPER1
G protein-coupled estrogen receptor 1
SEQ ID NOS: 6140-6146

GPHA2
Glycoprotein hormone alpha 2
SEQ ID NOS: 6147-6149

GPHB5
Glycoprotein hormone beta 5
SEQ ID NOS: 6150-6151

GPIHBP1
Glycosylphosphatidylinositol anchored high density
SEQ ID NO: 6152

lipoprotein binding protein 1

GPLD1
Glycosylphosphatidylinositol specific phospholipase
SEQ ID NO: 6153

D1

GPNMB
Glycoprotein (transmembrane) nmb
SEQ ID NOS: 6154-6156

GPR162
G protein-coupled receptor 162
SEQ ID NOS: 6157-6160

GPX3
Glutathione peroxidase 3
SEQ ID NOS: 6161-6168

GPX4
Glutathione peroxidase 4
SEQ ID NOS: 6169-6179

GPX5
Glutathione peroxidase 5
SEQ ID NOS: 6180-6181

GPX6
Glutathione peroxidase 6
SEQ ID NOS: 6182-6184

GPX7
Glutathione peroxidase 7
SEQ ID NO: 6185

GREM1
Gremlin 1, DAN family BMP antagonist
SEQ ID NOS: 6186-6188

GREM2
Gremlin 2, DAN family BMP antagonist
SEQ ID NO: 6189

GRHL3
Grainyhead-like transcription factor 3
SEQ ID NOS: 6190-6195

GRIA2
Glutamate receptor, ionotropic, AMPA 2
SEQ ID NOS: 6196-6207

GRIA3
Glutamate receptor, ionotropic, AMPA 3
SEQ ID NOS: 6208-6213

GRIA4
Glutamate receptor, ionotropic, AMPA 4
SEQ ID NOS: 6214-6225

GRIK2
Glutamate receptor, ionotropic, kainate 2
SEQ ID NOS: 6226-6234

GRIN2B
Glutamate receptor, ionotropic, N-methyl D-
SEQ ID NOS: 6235-6238

aspartate 2B

GRM2
Glutamate receptor, metabotropic 2
SEQ ID NOS: 6239-6242

GRM3
Glutamate receptor, metabotropic 3
SEQ ID NOS: 6243-6247

GRM5
Glutamate receptor, metabotropic 5
SEQ ID NOS: 6248-6252

GRN
Granulin
SEQ ID NOS: 6253-6268

GRP
Gastrin-releasing peptide
SEQ ID NOS: 6269-6273

DFNA5
Deafness, autosomal dominant 5
SEQ ID NOS: 6274-6282

GSG1
Germ cell associated 1
SEQ ID NOS: 6283-6291

GSN
Gelsolin
SEQ ID NOS: 6292-6300

GTDC1
Glycosyltransferase-like domain containing 1
SEQ ID NOS: 6301-6314

GTPBP10
GTP-binding protein 10 (putative)
SEQ ID NOS: 6315-6323

GUCA2A
Guanylate cyclase activator 2A (guanylin)
SEQ ID NO: 6324

GUCA2B
Guanylate cyclase activator 2B (uroguanylin)
SEQ ID NO: 6325

GUSB
Glucuronidase, beta
SEQ ID NOS: 6326-6330

GVQW1
GVQW motif containing 1
SEQ ID NO: 6331

GXYLT1
Glucoside xylosyltransferase 1
SEQ ID NOS: 6332-6333

GXYLT2
Glucoside xylosyltransferase 2
SEQ ID NOS: 6334-6336

GYPB
Glycophorin B (MNS blood group)
SEQ ID NOS: 6337-6345

GZMA
Granzyme A (granzyme 1, cytotoxic T-lymphocyte-
SEQ ID NO: 6346

associated serine esterase 3)

GZMB
Granzyme B (granzyme 2, cytotoxic T-lymphocyte-
SEQ ID NOS: 6347-6355

associated serine esterase 1)

GZMH
Granzyme H (cathepsin G-like 2, protein h-CCPX)
SEQ ID NOS: 6356-6358

GZMK
Granzyme K (granzyme 3; tryptase II)
SEQ ID NO: 6359

GZMM
Granzyme M (lymphocyte met-ase 1)
SEQ ID NOS: 6360-6361

H6PD
Hexose-6-phosphate dehydrogenase (glucose 1-
SEQ ID NOS: 6362-6363

dehydrogenase)

HABP2
Hyaluronan binding protein 2
SEQ ID NOS: 6364-6365

HADHB
Hydroxyacyl-CoA dehydrogenase/3-ketoacyl-CoA
SEQ ID NOS: 6366-6372

thiolase/enoyl-CoA hydratase (trifunctional protein),

beta subunit

HAMP
Hepcidin antimicrobial peptide
SEQ ID NOS: 6373-6374

HAPLN1
Hyaluronan and proteoglycan link protein 1
SEQ ID NOS: 6375-6381

HAPLN2
Hyaluronan and proteoglycan link protein 2
SEQ ID NOS: 6382-6383

HAPLN3
Hyaluronan and proteoglycan link protein 3
SEQ ID NOS: 6384-6387

HAPLN4
Hyaluronan and proteoglycan link protein 4
SEQ ID NO: 6388

HARS2
Histidyl-tRNA synthetase 2, mitochondrial
SEQ ID NOS: 6389-6404

HAVCR1
Hepatitis A virus cellular receptor 1
SEQ ID NOS: 6405-6409

HCCS
Holocytochrome c synthase
SEQ ID NOS: 6410-6412

HCRT
Hypocretin (orexin) neuropeptide precursor
SEQ ID NO: 6413

CECR5
Cat eye syndrome chromosome region, candidate 5
SEQ ID NOS: 6414-6416

HEATR5A
HEAT repeat containing 5A
SEQ ID NOS: 6417-6423

HEPH
Hephaestin
SEQ ID NOS: 6424-6431

HEXA
Hexosaminidase A (alpha polypeptide)
SEQ ID NOS: 6432-6441

HEXB
Hexosaminidase B (beta polypeptide)
SEQ ID NOS: 6442-6447

HFE2
Hemochromatosis type 2 (juvenile)
SEQ ID NOS: 6448-6454

HGF
Hepatocyte growth factor (hepapoietin A; scatter
SEQ ID NOS: 6455-6465

factor)

HGFAC
HGF activator
SEQ ID NOS: 6466-6467

HHIP
Hedgehog interacting protein
SEQ ID NOS: 6468-6469

HHIPL1
HHIP-like 1
SEQ ID NOS: 6470-6471

HHIPL2
HHIP-like 2
SEQ ID NO: 6472

HHLA1
HERV-H LTR-associating 1
SEQ ID NOS: 6473-6474

HHLA2
HERV-H LTR-associating 2
SEQ ID NOS: 6475-6485

HIBADH
3-hydroxyisobutyrate dehydrogenase
SEQ ID NOS: 6486-6488

HINT2
Histidine triad nucleotide binding protein 2
SEQ ID NO: 6489

HLA-A
Major histocompatibility complex, class I, A
SEQ ID NOS: 6490-6494

HLA-C
Major histocompatibility complex, class I, C
SEQ ID NOS: 6495-6499

HLA-DOA
Major histocompatibility complex, class II, DO alpha
SEQ ID NOS: 6500-6501

HLA-DPA1
Major histocompatibility complex, class II, DP
SEQ ID NOS: 6502-6505

alpha 1

HLA-DQA1
Major histocompatibility complex, class II, DQ
SEQ ID NOS: 6506-6511

alpha 1

HLA-DQB1
Major histocompatibility complex, class II, DQ beta 1
SEQ ID NOS: 6512-6517

HLA-DQB2
Major histocompatibility complex, class II, DQ beta 2
SEQ ID NOS: 6518-6521

HMCN1
Hemicentin 1
SEQ ID NOS: 6522-6523

HMCN2
Hemicentin 2
SEQ ID NOS: 6524-6527

HMGCL
3-hydroxymethyl-3-methylglutaryl-CoA lyase
SEQ ID NOS: 6528-6531

HMSD
Histocompatibility (minor) serpin domain containing
SEQ ID NOS: 6532-6533

HP
Haptoglobin
SEQ ID NOS: 6534-6547

HPR
Haptoglobin-related protein
SEQ ID NOS: 6548-6550

HPSE
Heparanase
SEQ ID NOS: 6551-6557

HPSE2
Heparanase 2 (inactive)
SEQ ID NOS: 6558-6563

HPX
Hemopexin
SEQ ID NOS: 6564-6565

HRC
Histidine rich calcium binding protein
SEQ ID NOS: 6566-6568

HRG
Histidine-rich glycoprotein
SEQ ID NO: 6569

HS2ST1
Heparan sulfate 2-O-sulfotransferase 1
SEQ ID NOS: 6570-6572

HS3ST1
Heparan sulfate (glucosamine) 3-O-
SEQ ID NOS: 6573-6575

sulfotransferase 1

HS6ST1
Heparan sulfate 6-O-sulfotransferase 1
SEQ ID NO: 6576

HS6ST3
Heparan sulfate 6-O-sulfotransferase 3
SEQ ID NOS: 6577-6578

HSD11B1L
Hydroxysteroid (11-beta) dehydrogenase 1-like
SEQ ID NOS: 6579-6597

HSD17811
Hydroxysteroid (17-beta) dehydrogenase 11
SEQ ID NOS: 6598-6599

HSD17B7
Hydroxysteroid (17-beta) dehydrogenase 7
SEQ ID NOS: 6600-6604

HSP90B1
Heat shock protein 90 kDa beta (Grp94), member 1
SEQ ID NOS: 6605-6610

HSPA13
Heat shock protein 70 kDa family, member 13
SEQ ID NO: 6611

HSPA5
Heat shock 70 kDa protein 5 (glucose-regulated
SEQ ID NO: 6612

protein, 78 kDa)

HSPG2
Heparan sulfate proteoglycan 2
SEQ ID NOS: 6613-6617

HTATIP2
HIV-1 Tat interactive protein 2, 30 kDa
SEQ ID NOS: 6618-6625

HTN1
Histatin 1
SEQ ID NOS: 6626-6628

HTN3
Histatin 3
SEQ ID NOS: 6629-6631

HTRA1
HtrA serine peptidase 1
SEQ ID NOS: 6632-6633

HTRA3
HtrA serine peptidase 3
SEQ ID NOS: 6634-6635

HTRA4
HtrA serine peptidase 4
SEQ ID NO: 6636

HYAL1
Hyaluronoglucosaminidase 1
SEQ ID NOS: 6637-6645

HYAL2
Hyaluronoglucosaminidase 2
SEQ ID NOS: 6646-6654

HYAL3
Hyaluronoglucosaminidase 3
SEQ ID NOS: 6655-6661

HYOU1
Hypoxia up-regulated 1
SEQ ID NOS: 6662-6676

IAPP
Islet amyloid polypeptide
SEQ ID NOS: 6677-6681

IBSP
Integrin-binding sialoprotein
SEQ ID NO: 6682

ICAM1
Intercellular adhesion molecule 1
SEQ ID NOS: 6683-6685

ICAM2
Intercellular adhesion molecule 2
SEQ ID NOS: 6686-6696

ICAM4
Intercellular adhesion molecule 4 (Landsteiner-
SEQ ID NOS: 6697-6699

Wiener blood group)

ID1
Inhibitor of DNA binding 1, dominant negative helix-
SEQ ID NOS: 6700-6701

loop-helix protein

IDE
Insulin-degrading enzyme
SEQ ID NOS: 6702-6705

IDNK
IdnK, gluconokinase homolog (E. coli)
SEQ ID NOS: 6706-6711

IDS
Iduronate 2-sulfatase
SEQ ID NOS: 6712-6717

IDUA
Iduronidase, alpha-L-
SEQ ID NOS: 6718-6723

IFI27L2
Interferon, alpha-inducible protein 27-like 2
SEQ ID NOS: 6724-6725

IFI30
Interferon, gamma-inducible protein 30
SEQ ID NOS: 6726-6727

IFNA1
Interferon, alpha 1
SEQ ID NO: 6728

IFNA10
Interferon, alpha 10
SEQ ID NO: 6729

IFNA13
Interferon, alpha 13
SEQ ID NOS: 6730-6731

IFNA14
Interferon, alpha 14
SEQ ID NO: 6732

IFNA16
Interferon, alpha 16
SEQ ID NO: 6733

IFNA17
Interferon, alpha 17
SEQ ID NO: 6734

IFNA2
Interferon, alpha 2
SEQ ID NO: 6735

IFNA21
Interferon, alpha 21
SEQ ID NO: 6736

IFNA4
Interferon, alpha 4
SEQ ID NO: 6737

IFNA5
Interferon, alpha 5
SEQ ID NO: 6738

IFNA6
Interferon, alpha 6
SEQ ID NOS: 6739-6740

IFNA7
Interferon, alpha 7
SEQ ID NO: 6741

IFNA8
Interferon, alpha 8
SEQ ID NO: 6742

IFNAR1
Interferon (alpha, beta and omega) receptor 1
SEQ ID NOS: 6743-6744

IFNB1
Interferon, beta 1, fibroblast
SEQ ID NO: 6745

IFNE
Interferon, epsilon
SEQ ID NO: 6746

IFNG
Interferon, gamma
SEQ ID NO: 6747

IFNGR1
Interferon gamma receptor 1
SEQ ID NOS: 6748-6758

IFNL1
Interferon, lambda 1
SEQ ID NO: 6759

IFNL2
Interferon, lambda 2
SEQ ID NO: 6760

IFNL3
Interferon, lambda 3
SEQ ID NOS: 6761-6762

IFNLR1
Interferon, lambda receptor 1
SEQ ID NOS: 6763-6767

IFNW1
Interferon, omega 1
SEQ ID NO: 6768

IGF1
Insulin-like growth factor 1 (somatomedin C)
SEQ ID NOS: 6769-6774

IGF2
Insulin-like growth factor 2
SEQ ID NOS: 6775-6782

IGFALS
Insulin-like growth factor binding protein, acid labile
SEQ ID NOS: 6783-6785

subunit

IGFBP1
Insulin-like growth factor binding protein 1
SEQ ID NOS: 6786-6788

IGFBP2
Insulin-like growth factor binding protein 2, 36 kDa
SEQ ID NOS: 6789-6792

IGFBP3
Insulin-like growth factor binding protein 3
SEQ ID NOS: 6793-6800

IGFBP4
Insulin-like growth factor binding protein 4
SEQ ID NO: 6801

IGFBP5
Insulin-like growth factor binding protein 5
SEQ ID NOS: 6802-6803

IGFBP6
Insulin-like growth factor binding protein 6
SEQ ID NOS: 6804-6806

IGFBP7
Insulin-like growth factor binding protein 7
SEQ ID NOS: 6807-6808

IGFBPL1
Insulin-like growth factor binding protein-like 1
SEQ ID NO: 6809

IGFL1
IGF-like family member 1
SEQ ID NO: 6810

IGFL2
IGF-like family member 2
SEQ ID NOS: 6811-6813

IGFL3
IGF-like family member 3
SEQ ID NO: 6814

IGFLR1
IGF-like family receptor 1
SEQ ID NOS: 6815-6823

IGIP
IgA-inducing protein
SEQ ID NO: 6824

IGLON5
IgLON family member 5
SEQ ID NO: 6825

IGSF1
Immunoglobulin superfamily, member 1
SEQ ID NOS: 6826-6831

IGSF10
Immunoglobulin superfamily, member 10
SEQ ID NOS: 6832-6833

IGSF11
Immunoglobulin superfamily, member 11
SEQ ID NOS: 6834-6841

IGSF21
Immunoglobin superfamily, member 21
SEQ ID NO: 6842

IGSF8
Immunoglobulin superfamily, member 8
SEQ ID NOS: 6843-6846

IGSF9
Immunoglobulin superfamily, member 9
SEQ ID NOS: 6847-6849

IHH
Indian hedgehog
SEQ ID NO: 6850

IL10
Interleukin 10
SEQ ID NOS: 6851-6852

IL11
Interleukin 11
SEQ ID NOS: 6853-6856

IL11RA
Interleukin 11 receptor, alpha
SEQ ID NOS: 6857-6867

IL12B
Interleukin 12B
SEQ ID NO: 6868

IL12RB1
Interleukin 12 receptor, beta 1
SEQ ID NOS: 6869-6874

IL12RB2
Interleukin 12 receptor, beta 2
SEQ ID NOS: 6875-6879

IL13
Interleukin 13
SEQ ID NOS: 6880-6881

IL13RA1
Interleukin 13 receptor, alpha 1
SEQ ID NOS: 6882-6883

IL15RA
Interleukin 15 receptor, alpha
SEQ ID NOS: 6884-6901

IL17A
Interleukin 17A
SEQ ID NO: 6902

IL17B
Interleukin 17B
SEQ ID NO: 6903

IL17C
Interleukin 17C
SEQ ID NO: 6904

IL17D
Interleukin 17D
SEQ ID NOS: 6905-6907

IL17F
Interleukin 17F
SEQ ID NO: 6908

IL17RA
Interleukin 17 receptor A
SEQ ID NOS: 6909-6910

IL17RC
Interleukin 17 receptor C
SEQ ID NOS: 6911-6926

IL17RE
Interleukin 17 receptor E
SEQ ID NOS: 6927-6933

IL18BP
Interleukin 18 binding protein
SEQ ID NOS: 6934-6944

IL18R1
Interleukin 18 receptor 1
SEQ ID NOS: 6945-6948

IL18RAP
Interleukin 18 receptor accessory protein
SEQ ID NOS: 6949-6951

IL19
Interleukin 19
SEQ ID NOS: 6952-6954

IL1R1
Interleukin 1 receptor, type I
SEQ ID NOS: 6955-6967

IL1R2
Interleukin 1 receptor, type II
SEQ ID NOS: 6968-6971

IL1RAP
Interleukin 1 receptor accessory protein
SEQ ID NOS: 6972-6985

IL1RL1
Interleukin 1 receptor-like 1
SEQ ID NOS: 6986-6991

IL1RL2
Interleukin 1 receptor-like 2
SEQ ID NOS: 6992-6994

IL1RN
Interleukin 1 receptor antagonist
SEQ ID NOS: 6995-6999

IL2
Interleukin 2
SEQ ID NO: 7000

IL20
Interleukin 20
SEQ ID NOS: 7001-7003

IL20RA
Interleukin 20 receptor, alpha
SEQ ID NOS: 7004-7010

IL21
Interleukin 21
SEQ ID NOS: 7011-7012

IL22
Interleukin 22
SEQ ID NOS: 7013-7014

IL22RA2
Interleukin 22 receptor, alpha 2
SEQ ID NOS: 7015-7017

IL23A
Interleukin 23, alpha subunit p19
SEQ ID NO: 7018

IL24
Interleukin 24
SEQ ID NOS: 7019-7024

IL25
Interleukin 25
SEQ ID NOS: 7025-7026

IL26
Interleukin 26
SEQ ID NO: 7027

IL27
Interleukin 27
SEQ ID NOS: 7028-7029

IL2RB
Interleukin 2 receptor, beta
SEQ ID NOS: 7030-7034

IL3
Interleukin 3
SEQ ID NO: 7035

IL31
Interleukin 31
SEQ ID NO: 7036

IL31RA
Interleukin 31 receptor A
SEQ ID NOS: 7037-7044

IL32
Interleukin 32
SEQ ID NOS: 7045-7074

IL34
Interleukin 34
SEQ ID NOS: 7075-7078

IL3RA
Interleukin 3 receptor, alpha (low affinity)
SEQ ID NOS: 7079-7081

IL4
Interleukin 4
SEQ ID NOS: 7082-7084

IL4I1
Interleukin 4 induced 1
SEQ ID NOS: 7085-7092

IL4R
Interleukin 4 receptor
SEQ ID NOS: 7093-7106

IL5
Interleukin 5
SEQ ID NOS: 7107-7108

IL5RA
Interleukin 5 receptor, alpha
SEQ ID NOS: 7109-7118

IL6
Interleukin 6
SEQ ID NOS: 7119-7125

IL6R
Interleukin 6 receptor
SEQ ID NOS: 7126-7131

IL6ST
Interleukin 6 signal transducer
SEQ ID NOS: 7132-7141

IL7
Interleukin 7
SEQ ID NOS: 7142-7149

IL7R
Interleukin 7 receptor
SEQ ID NOS: 7150-7156

IL9
Interleukin 9
SEQ ID NO: 7157

ILDR1
Immunoglobulin-like domain containing receptor 1
SEQ ID NOS: 7158-7162

ILDR2
Immunoglobulin-like domain containing receptor 2
SEQ ID NOS: 7163-7169

IMP4
IMP4, U3 small nucleolar ribonucleoprotein
SEQ ID NOS: 7170-7175

IMPG1
Interphotoreceptor matrix proteoglycan 1
SEQ ID NOS: 7176-7179

INHA
Inhibin, alpha
SEQ ID NO: 7180

INHBA
Inhibin, beta A
SEQ ID NOS: 7181-7183

INHBB
Inhibin, beta B
SEQ ID NO: 7184

INHBC
Inhibin, beta C
SEQ ID NO: 7185

INHBE
Inhibin, beta E
SEQ ID NOS: 7186-7187

INPP5A
Inositol polyphosphate-5-phosphatase A
SEQ ID NOS: 7188-7192

INS
Insulin
SEQ ID NOS: 7193-7197

INS-IGF2
INS-IGF2 readthrough
SEQ ID NOS: 7198-7199

INSL3
Insulin-like 3 (Leydig cell)
SEQ ID NOS: 7200-7202

INSL4
Insulin-like 4 (placenta)
SEQ ID NO: 7203

INSL5
Insulin-like 5
SEQ ID NO: 7204

INSL6
Insulin-like 6
SEQ ID NO: 7205

INTS3
Integrator complex subunit 3
SEQ ID NOS: 7206-7211

IPO11
Importin 11
SEQ ID NOS: 7212-7220

IPO9
Importin 9
SEQ ID NOS: 7221-7222

IQCF6
IQ motif containing F6
SEQ ID NOS: 7223-7224

IRAK3
Interleukin-1 receptor-associated kinase 3
SEQ ID NOS: 7225-7227

IRS4
Insulin receptor substrate 4
SEQ ID NO: 7228

ISLR
Immunoglobulin superfamily containing leucine-rich
SEQ ID NOS: 7229-7232

repeat

ISLR2
Immunoglobulin superfamily containing leucine-rich
SEQ ID NOS: 7233-7242

repeat 2

ISM1
Isthmin 1, angiogenesis inhibitor
SEQ ID NO: 7243

ISM2
Isthmin 2
SEQ ID NOS: 7244-7249

ITGA4
Integrin, alpha 4 (antigen CD49D, alpha 4 subunit of
SEQ ID NOS: 7250-7252

VLA-4 receptor)

ITGA9
Integrin, alpha 9
SEQ ID NOS: 7253-7255

ITGAL
Integrin, alpha L (antigen CD11A (p180), lymphocyte
SEQ ID NOS: 7256-7265

function-associated antigen 1; alpha polypeptide)

ITGAX
Integrin, alpha X (complement component 3
SEQ ID NOS: 7266-7268

receptor 4 subunit)

ITGB1
Integrin, beta 1 (fibronectin receptor, beta
SEQ ID NOS: 7269-7284

polypeptide, antigen CD29 includes MDF2, MSK12)

ITGB2
Integrin, beta 2 (complement component 3 receptor
SEQ ID NOS: 7285-7301

3 and 4 subunit)

ITGB3
Integrin, beta 3 (platelet glycoprotein IIIa, antigen
SEQ ID NOS: 7302-7304

CD61)

ITGB7
Integrin, beta 7
SEQ ID NOS: 7305-7312

ITGBL1
Integrin, beta-like 1 (with EGF-like repeat domains)
SEQ ID NOS: 7313-7318

ITIH1
Inter-alpha-trypsin inhibitor heavy chain 1
SEQ ID NOS: 7319-7324

ITIH2
Inter-alpha-trypsin inhibitor heavy chain 2
SEQ ID NOS: 7325-7327

ITIH3
Inter-alpha-trypsin inhibitor heavy chain 3
SEQ ID NOS: 7328-7330

ITIH4
Inter-alpha-trypsin inhibitor heavy chain family,
SEQ ID NOS: 7331-7334

member 4

ITIH5
Inter-alpha-trypsin inhibitor heavy chain family,
SEQ ID NOS: 7335-7338

member 5

ITIH6
Inter-alpha-trypsin inhibitor heavy chain family,
SEQ ID NO: 7339

member 6

ITLN1
Intelectin 1 (galactofuranose binding)
SEQ ID NO: 7340

ITLN2
Intelectin 2
SEQ ID NO: 7341

IZUMO1R
IZUMO1 receptor, JUNO
SEQ ID NOS: 7342-7343

IZUMO4
IZUMO family member 4
SEQ ID NOS: 7344-7350

AMICA1
Adhesion molecule, interacts with CXADR antigen 1
SEQ ID NOS: 7351-7359

JCHAIN
Joining chain of multimeric IgA and IgM
SEQ ID NOS: 7360-7365

JMJD8
Jumonji domain containing 8
SEQ ID NOS: 7366-7370

JSRP1
Junctional sarcoplasmic reticulum protein 1
SEQ ID NO: 7371

KANSL2
KAT8 regulatory NSL complex subunit 2
SEQ ID NOS: 7372-7382

KAZALD1
Kazal-type serine peptidase inhibitor domain 1
SEQ ID NO: 7383

KCNIP3
Kv channel interacting protein 3, calsenilin
SEQ ID NOS: 7384-7386

KCNK7
Potassium channel, two pore domain subfamily K,
SEQ ID NOS: 7387-7392

member 7

KCNN4
Potassium channel, calcium activated
SEQ ID NOS: 7393-7398

intermediate/small conductance subfamily N alpha,

member 4

KCNU1
Potassium channel, subfamily U, member 1
SEQ ID NOS: 7399-7403

KCP
Kielin/chordin-like protein
SEQ ID NOS: 7404-7407

KDELC1
KDEL (Lys-Asp-Glu-Leu) containing 1
SEQ ID NO: 7408

KDELC2
KDEL (Lys-Asp-Glu-Leu) containing 2
SEQ ID NOS: 7409-7412

KDM1A
Lysine (K)-specific demethylase 1A
SEQ ID NOS: 7413-7416

KDM3B
Lysine (K)-specific demethylase 3B
SEQ ID NOS: 7417-7420

KDM6A
Lysine (K)-specific demethylase 6A
SEQ ID NOS: 7421-7430

KDM7A
Lysine (K)-specific demethylase 7A
SEQ ID NOS: 7431-7432

KDSR
3-ketodihydrosphingosine reductase
SEQ ID NOS: 7433-7439

KERA
Keratocan
SEQ ID NO: 7440

KIAA0100
KIAA0100
SEQ ID NOS: 7441-7446

KIAA0319
KIAA0319
SEQ ID NOS: 7447-7452

KIAA1324
KIAA1324
SEQ ID NOS: 7453-7461

KIFC2
Kinesin family member C2
SEQ ID NOS: 7462-7464

KIR2DL4
Killer cell immunoglobulin-like receptor, two
SEQ ID NOS: 7465-7471

domains, long cytoplasmic tail, 4

KIR3DX1
Killer cell immunoglobulin-like receptor, three
SEQ ID NOS: 7472-7476

domains, X1

KIRREL2
Kin of IRRE like 2 (Drosophila)
SEQ ID NOS: 7477-7481

KISS1
KiSS-1 metastasis-suppressor
SEQ ID NOS: 7482-7483

KLHL11
Kelch-like family member 11
SEQ ID NO: 7484

KLHL22
Kelch-like family member 22
SEQ ID NOS: 7485-7491

KLK1
Kallikrein 1
SEQ ID NOS: 7492-7493

KLK10
Kallikrein-related peptidase 10
SEQ ID NOS: 7494-7498

KLK11
Kallikrein-related peptidase 11
SEQ ID NOS: 7499-7507

KLK12
Kallikrein-related peptidase 12
SEQ ID NOS: 7508-7514

KLK13
Kallikrein-related peptidase 13
SEQ ID NOS: 7515-7523

KLK14
Kallikrein-related peptidase 14
SEQ ID NOS: 7524-7525

KLK15
Kallikrein-related peptidase 15
SEQ ID NOS: 7526-7530

KLK2
Kallikrein-related peptidase 2
SEQ ID NOS: 7531-7543

KLK3
Kallikrein-related peptidase 3
SEQ ID NOS: 7544-7555

KLK4
Kallikrein-related peptidase 4
SEQ ID NOS: 7556-7560

KLK5
Kallikrein-related peptidase 5
SEQ ID NOS: 7561-7564

KLK6
Kallikrein-related peptidase 6
SEQ ID NOS: 7565-7571

KLK7
Kallikrein-related peptidase 7
SEQ ID NOS: 7572-7576

KLK8
Kallikrein-related peptidase 8
SEQ ID NOS: 7577-7584

KLK9
Kallikrein-related peptidase 9
SEQ ID NOS: 7585-7586

KLKB1
Kallikrein B, plasma (Fletcher factor) 1
SEQ ID NOS: 7587-7591

SETD8
SET domain containing (lysine methyltransferase) 8
SEQ ID NOS: 7592-7595

KNDC1
Kinase non-catalytic C-lobe domain (KIND)
SEQ ID NOS: 7596-7597

containing 1

KNG1
Kininogen 1
SEQ ID NOS: 7598-7602

KRBA2
KRAB-A domain containing 2
SEQ ID NOS: 7603-7606

KREMEN2
Kringle containing transmembrane protein 2
SEQ ID NOS: 7607-7612

KRTDAP
Keratinocyte differentiation-associated protein
SEQ ID NOS: 7613-7614

L1CAM
L1 cell adhesion molecule
SEQ ID NOS: 7615-7624

L3MBTL2
L(3)mbt-like 2 (Drosophila)
SEQ ID NOS: 7625-7629

LACRT
Lacritin
SEQ ID NOS: 7630-7632

LACTB
Lactamase, beta
SEQ ID NOS: 7633-7635

LAG3
Lymphocyte-activation gene 3
SEQ ID NOS: 7636-7637

LAIR2
Leukocyte-associated immunoglobulin-like
SEQ ID NOS: 7638-7641

receptor 2

LALBA
Lactalbumin, alpha-
SEQ ID NOS: 7642-7643

LAMA1
Laminin, alpha 1
SEQ ID NOS: 7644-7645

LAMA2
Laminin, alpha 2
SEQ ID NOS: 7646-7649

LAMA3
Laminin, alpha 3
SEQ ID NOS: 7650-7659

LAMA4
Laminin, alpha 4
SEQ ID NOS: 7660-7674

LAMAS
Laminin, alpha 5
SEQ ID NOS: 7675-7677

LAMB1
Laminin, beta 1
SEQ ID NOS: 7678-7682

LAMB2
Laminin, beta 2 (laminin S)
SEQ ID NOS: 7683-7685

LAMB3
Laminin, beta 3
SEQ ID NOS: 7686-7690

LAMB4
Laminin, beta 4
SEQ ID NOS: 7691-7694

LAMC1
Laminin, gamma 1 (formerly LAMB2)
SEQ ID NOS: 7695-7696

LAMC2
Laminin, gamma 2
SEQ ID NOS: 7697-7698

LAMC3
Laminin, gamma 3
SEQ ID NOS: 7699-7700

LAMP3
Lysosomal-associated membrane protein 3
SEQ ID NOS: 7701-7704

GYLTL1B
Glycosyltransferase-like 1B
SEQ ID NOS: 7705-7710

LAT
Linker for activation of T cells
SEQ ID NOS: 7711-7720

LAT2
Linker for activation of T cells family, member 2
SEQ ID NOS: 7721-7729

LBP
Lipopolysaccharide binding protein
SEQ ID NO: 7730

LCAT
Lecithin-cholesterol acyltransferase
SEQ ID NOS: 7731-7737

LCN1
Lipocalin 1
SEQ ID NOS: 7738-7739

LCN10
Lipocalin 10
SEQ ID NOS: 7740-7745

LCN12
Lipocalin 12
SEQ ID NOS: 7746-7748

LCN15
Lipocalin 15
SEQ ID NO: 7749

LCN2
Lipocalin 2
SEQ ID NOS: 7750-7752

LCN6
Lipocalin 6
SEQ ID NOS: 7753-7754

LCN8
Lipocalin 8
SEQ ID NOS: 7755-7756

LCN9
Lipocalin 9
SEQ ID NOS: 7757-7758

LCORL
Ligand dependent nuclear receptor corepressor-like
SEQ ID NOS: 7759-7764

LDLR
Low density lipoprotein receptor
SEQ ID NOS: 7765-7773

LDLRAD2
Low density lipoprotein receptor class A domain
SEQ ID NOS: 7774-7775

containing 2

LEAP2
Liver expressed antimicrobial peptide 2
SEQ ID NO: 7776

LECT2
Leukocyte cell-derived chemotaxin 2
SEQ ID NOS: 7777-7780

LEFTY1
Left-right determination factor 1
SEQ ID NOS: 7781-7782

LEFTY2
Left-right determination factor 2
SEQ ID NOS: 7783-7784

LEP
Leptin
SEQ ID NO: 7785

LFNG
LFNG O-fucosylpeptide 3-beta-N-
SEQ ID NOS: 7786-7791

acetylglucosaminyltransferase

LGALS3BP
Lectin, galactoside-binding, soluble, 3 binding
SEQ ID NOS: 7792-7806

protein

LGI1
Leucine-rich, glioma inactivated 1
SEQ ID NOS: 7807-7825

LGI2
Leucine-rich repeat LGI family, member 2
SEQ ID NOS: 7826-7827

LGI3
Leucine-rich repeat LGI family, member 3
SEQ ID NOS: 7828-7831

LGI4
Leucine-rich repeat LGI family, member 4
SEQ ID NOS: 7832-7835

LGMN
Legumain
SEQ ID NOS: 7836-7849

LGR4
Leucine-rich repeat containing G protein-coupled
SEQ ID NOS: 7850-7852

receptor 4

LHB
Luteinizing hormone beta polypeptide
SEQ ID NO: 7853

LHCGR
Luteinizing hormone/choriogonadotropin receptor
SEQ ID NOS: 7854-7858

LIF
Leukemia inhibitory factor
SEQ ID NOS: 7859-7860

LIFR
Leukemia inhibitory factor receptor alpha
SEQ ID NOS: 7861-7865

LILRA1
Leukocyte immunoglobulin-like receptor, subfamily
SEQ ID NOS: 7866-7867

A (with TM domain), member 1

LILRA2
Leukocyte immunoglobulin-like receptor, subfamily
SEQ ID NOS: 7868-7874

A (with TM domain), member 2

LILRB3
Leukocyte immunoglobulin-like receptor, subfamily
SEQ ID NOS: 7875-7879

B (with TM and ITIM domains), member 3

LIME1
Lck interacting transmembrane adaptor 1
SEQ ID NOS: 7880-7885

LINGO1
Leucine rich repeat and Ig domain containing 1
SEQ ID NOS: 7886-7896

LIPA
Lipase A, lysosomal acid, cholesterol esterase
SEQ ID NOS: 7897-7901

LIPC
Lipase, hepatic
SEQ ID NOS: 7902-7905

LIPF
Lipase, gastric
SEQ ID NOS: 7906-7909

LIPG
Lipase, endothelial
SEQ ID NOS: 7910-7915

LIPH
Lipase, member H
SEQ ID NOS: 7916-7920

LIPK
Lipase, family member K
SEQ ID NO: 7921

LIPM
Lipase, family member M
SEQ ID NOS: 7922-7923

LIPN
Lipase, family member N
SEQ ID NO: 7924

LMAN2
Lectin, mannose-binding 2
SEQ ID NOS: 7925-7929

LMNTD1
Lamin tail domain containing 1
SEQ ID NOS: 7930-7940

LNX1
Ligand of numb-protein X 1, E3 ubiquitin protein
SEQ ID NOS: 7941-7947

ligase

LOX
Lysyl oxidase
SEQ ID NOS: 7948-7950

LOXL1
Lysyl oxidase-like 1
SEQ ID NOS: 7951-7952

LOXL2
Lysyl oxidase-like 2
SEQ ID NOS: 7953-7961

LOXL3
Lysyl oxidase-like 3
SEQ ID NOS: 7962-7968

LOXL4
Lysyl oxidase-like 4
SEQ ID NO: 7969

LPA
Lipoprotein, Lp(a)
SEQ ID NOS: 7970-7972

LPL
Lipoprotein lipase
SEQ ID NOS: 7973-7977

LPO
Lactoperoxidase
SEQ ID NOS: 7978-7984

LRAT
Lecithin retinol acyltransferase
SEQ ID NOS: 7985-7987

(phosphatidylcholine-retinol O-acyltransferase)

LRCH3
Leucine-rich repeats and calponin homology (CH)
SEQ ID NOS: 7988-7996

domain containing 3

LRCOL1
Leucine rich colipase-like 1
SEQ ID NOS: 7997-8000

LRFN4
Leucine rich repeat and fibronectin type III domain
SEQ ID NOS: 8001-8002

containing 4

LRFN5
Leucine rich repeat and fibronectin type III domain
SEQ ID NOS: 8003-8005

containing 5

LRG1
Leucine-rich alpha-2-glycoprotein 1
SEQ ID NO: 8006

LRP1
Low density lipoprotein receptor-related protein 1
SEQ ID NOS: 8007-8012

LRP11
Low density lipoprotein receptor-related protein 11
SEQ ID NOS: 8013-8014

LRP1B
Low density lipoprotein receptor-related protein 1B
SEQ ID NOS: 8015-8018

LRP2
Low density lipoprotein receptor-related protein 2
SEQ ID NOS: 8019-8020

LRP4
Low density lipoprotein receptor-related protein 4
SEQ ID NOS: 8021-8022

LRPAP1
Low density lipoprotein receptor-related protein
SEQ ID NOS: 8023-8024

associated protein 1

LRRC17
Leucine rich repeat containing 17
SEQ ID NOS: 8025-8027

LRRC32
Leucine rich repeat containing 32
SEQ ID NOS: 8028-8031

LRRC3B
Leucine rich repeat containing 3B
SEQ ID NOS: 8032-8036

LRRC4B
Leucine rich repeat containing 4B
SEQ ID NOS: 8037-8039

LRRC70
Leucine rich repeat containing 70
SEQ ID NOS: 8040-8041

LRRN3
Leucine rich repeat neuronal 3
SEQ ID NOS: 8042-8045

LRRTM1
Leucine rich repeat transmembrane neuronal 1
SEQ ID NOS: 8046-8052

LRRTM2
Leucine rich repeat transmembrane neuronal 2
SEQ ID NOS: 8053-8055

LRRTM4
Leucine rich repeat transmembrane neuronal 4
SEQ ID NOS: 8056-8061

LRTM2
Leucine-rich repeats and transmembrane domains 2
SEQ ID NOS: 8062-8066

LSR
Lipolysis stimulated lipoprotein receptor
SEQ ID NOS: 8067-8077

LST1
Leukocyte specific transcript 1
SEQ ID NOS: 8078-8095

LTA
Lymphotoxin alpha
SEQ ID NOS: 8096-8097

LTBP1
Latent transforming growth factor beta binding
SEQ ID NOS: 8098-8107

protein 1

LTBP2
Latent transforming growth factor beta binding
SEQ ID NOS: 8108-8111

protein 2

LTBP3
Latent transforming growth factor beta binding
SEQ ID NOS: 8112-8124

protein 3

LTBP4
Latent transforming growth factor beta binding
SEQ ID NOS: 8125-8140

protein 4

LTBR
Lymphotoxin beta receptor (TNFR superfamily,
SEQ ID NOS: 8141-8146

member 3)

LTF
Lactotransferrin
SEQ ID NOS: 8147-8151

LTK
Leukocyte receptor tyrosine kinase
SEQ ID NOS: 8152-8155

LUM
Lumican
SEQ ID NO: 8156

LUZP2
Leucine zipper protein 2
SEQ ID NOS: 8157-8160

LVRN
Laeverin
SEQ ID NOS: 8161-8166

LY6E
Lymphocyte antigen 6 complex, locus E
SEQ ID NOS: 8167-8180

LY6G5B
Lymphocyte antigen 6 complex, locus G5B
SEQ ID NOS: 8181-8182

LY6G6D
Lymphocyte antigen 6 complex, locus G6D
SEQ ID NOS: 8183-8184

LY6G6E
Lymphocyte antigen 6 complex, locus G6E
SEQ ID NOS: 8185-8188

(pseudogene)

LY6H
Lymphocyte antigen 6 complex, locus H
SEQ ID NOS: 8189-8192

LY6K
Lymphocyte antigen 6 complex, locus K
SEQ ID NOS: 8193-8196

RP11-

SEQ ID NO: 8197

520P18.5

LY86
Lymphocyte antigen 86
SEQ ID NOS: 8198-8199

LY96
Lymphocyte antigen 96
SEQ ID NOS: 8200-8201

LYG1
Lysozyme G-like 1
SEQ ID NOS: 8202-8203

LYG2
Lysozyme G-like 2
SEQ ID NOS: 8204-8209

LYNX1
Ly6/neurotoxin 1
SEQ ID NOS: 8210-8214

LYPD1
LY6/PLAUR domain containing 1
SEQ ID NOS: 8215-8217

LYPD2
LY6/PLAUR domain containing 2
SEQ ID NO: 8218

LYPD4
LY6/PLAUR domain containing 4
SEQ ID NOS: 8219-8221

LYPD6
LY6/PLAUR domain containing 6
SEQ ID NOS: 8222-8226

LYPD6B
LY6/PLAUR domain containing 6B
SEQ ID NOS: 8227-8233

LYPD8
LY6/PLAUR domain containing 8
SEQ ID NOS: 8234-8235

LYZ
Lysozyme
SEQ ID NOS: 8236-8238

LYZL4
Lysozyme-like 4
SEQ ID NOS: 8239-8240

LYZL6
Lysozyme-like 6
SEQ ID NOS: 8241-8243

M6PR
Mannose-6-phosphate receptor (cation dependent)
SEQ ID NOS: 8244-8254

MAD1L1
MAD1 mitotic arrest deficient-like 1 (yeast)
SEQ ID NOS: 8255-8267

MAG
Myelin associated glycoprotein
SEQ ID NOS: 8268-8273

MAGT1
Magnesium transporter 1
SEQ ID NOS: 8274-8277

MALSU1
Mitochondrial assembly of ribosomal large subunit 1
SEQ ID NO: 8278

MAMDC2
MAM domain containing 2
SEQ ID NO: 8279

MAN2B1
Mannosidase, alpha, class 2B, member 1
SEQ ID NOS: 8280-8285

MAN2B2
Mannosidase, alpha, class 2B, member 2
SEQ ID NOS: 8286-8288

MANBA
Mannosidase, beta A, lysosomal
SEQ ID NOS: 8289-8302

MANEAL
Mannosidase, endo-alpha-like
SEQ ID NOS: 8303-8307

MANF
Mesencephalic astrocyte-derived neurotrophic
SEQ ID NOS: 8308-8309

factor

MANSC1
MANSC domain containing 1
SEQ ID NOS: 8310-8313

MAP3K9
Mitogen-activated protein kinase 9
SEQ ID NOS: 8314-8319

MASP1
Mannan-binding lectin serine peptidase 1 (C4/C2
SEQ ID NOS: 8320-8327

activating component of Ra-reactive factor)

MASP2
Mannan-binding lectin serine peptidase 2
SEQ ID NOS: 8328-8329

MATN1
Matrilin 1, cartilage matrix protein
SEQ ID NO: 8330

MATN2
Matrilin 2
SEQ ID NOS: 8331-8343

MATN3
Matrilin 3
SEQ ID NOS: 8344-8345

MATN4
Matrilin 4
SEQ ID NOS: 8346-8350

MATR3
Matrin 3
SEQ ID NOS: 8351-8378

MAU2
MAU2 sister chromatid cohesion factor
SEQ ID NOS: 8379-8381

MAZ
MYC-associated zinc finger protein (purine-binding
SEQ ID NOS: 8382-8396

transcription factor)

MBD6
Methyl-CpG binding domain protein 6
SEQ ID NOS: 8397-8408

MBL2
Mannose-binding lectin (protein C) 2, soluble
SEQ ID NO: 8409

MBNL1
Muscleblind-like splicing regulator 1
SEQ ID NOS: 8410-8428

MCCC1
Methylcrotonoyl-CoA carboxylase 1 (alpha)
SEQ ID NOS: 8429-8440

MCCD1
Mitochondrial coiled-coil domain 1
SEQ ID NO: 8441

MCEE
Methylmalonyl CoA epimerase
SEQ ID NOS: 8442-8445

MCF2L
MCF.2 cell line derived transforming sequence-like
SEQ ID NOS: 8446-8467

MCFD2
Multiple coagulation factor deficiency 2
SEQ ID NOS: 8468-8479

MDFIC
MyoD family inhibitor domain containing
SEQ ID NOS: 8480-8487

MDGA1
MAM domain containing
SEQ ID NOS: 8488-8493

glycosylphosphatidylinositol anchor 1

MDK
Midkine (neurite growth-promoting factor 2)
SEQ ID NOS: 8494-8503

MED20
Mediator complex subunit 20
SEQ ID NOS: 8504-8508

MEGF10
Multiple EGF-like-domains 10
SEQ ID NOS: 8509-8512

MEGF6
Multiple EGF-like-domains 6
SEQ ID NOS: 8513-8516

MEI1
Meiotic double-stranded break formation protein 1
SEQ ID NOS: 8517-8520

MEI4
Meiotic double-stranded break formation protein 4
SEQ ID NO: 8521

MEIS1
Meis homeobox 1
SEQ ID NOS: 8522-8527

MEIS3
Meis homeobox 3
SEQ ID NOS: 8528-8537

MFI2
Antigen p97 (melanoma associated) identified by
SEQ ID NOS: 8538-8540

monoclonal antibodies 133.2 and 96.5

MEPE
Matrix extracellular phosphoglycoprotein
SEQ ID NOS: 8541-8547

MESDC2
Mesoderm development candidate 2
SEQ ID NOS: 8548-8552

MEST
Mesoderm specific transcript
SEQ ID NOS: 8553-8566

MET
MET proto-oncogene, receptor tyrosine kinase
SEQ ID NOS: 8567-8572

METRN
Meteorin, glial cell differentiation regulator
SEQ ID NOS: 8573-8577

METRNL
Meteorin, glial cell differentiation regulator-like
SEQ ID NOS: 8578-8581

METTL17
Methyltransferase like 17
SEQ ID NOS: 8582-8592

METTL24
Methyltransferase like 24
SEQ ID NO: 8593

METTL7B
Methyltransferase like 7B
SEQ ID NOS: 8594-8595

METTL9
Methyltransferase like 9
SEQ ID NOS: 8596-8604

MEX3C
Mex-3 RNA binding family member C
SEQ ID NOS: 8605-8607

MFAP2
Microfibrillar-associated protein 2
SEQ ID NOS: 8608-8609

MFAP3
Microfibrillar-associated protein 3
SEQ ID NOS: 8610-8614

MFAP3L
Microfibrillar-associated protein 3-like
SEQ ID NOS: 8615-8624

MFAP4
Microfibrillar-associated protein 4
SEQ ID NOS: 8625-8627

MFAP5
Microfibrillar associated protein 5
SEQ ID NOS: 8628-8638

MFGE8
Milk fat globule-EGF factor 8 protein
SEQ ID NOS: 8639-8645

MFNG
MFNG O-fucosylpeptide 3-beta-N-
SEQ ID NOS: 8646-8653

acetylglucosaminyltransferase

MGA
MGA, MAX dimerization protein
SEQ ID NOS: 8654-8662

MGAT2
Mannosyl (alpha-1,6-)-glycoprotein beta-1,2-N-
SEQ ID NO: 8663

acetylglucosaminyltransferase

MGAT3
Mannosyl (beta-1,4-)-glycoprotein beta-1,4-N-
SEQ ID NOS: 8664-8666

acetylglucosaminyltransferase

MGAT4A
Mannosyl (alpha-1,3-)-glycoprotein beta-1,4-N-
SEQ ID NOS: 8667-8671

acetylglucosaminyltransferase, isozyme A

MGAT4B
Mannosyl (alpha-1,3-)-glycoprotein beta-1,4-N-
SEQ ID NOS: 8672-8682

acetylglucosaminyltransferase, isozyme B

MGAT4D
MGAT4 family, member D
SEQ ID NOS: 8683-8688

MGLL
Monoglyceride lipase
SEQ ID NOS: 8689-8698

MGP
Matrix Gla protein
SEQ ID NOS: 8699-8701

MGST2
Microsomal glutathione S-transferase 2
SEQ ID NOS: 8702-8705

MIA
Melanoma inhibitory activity
SEQ ID NOS: 8706-8711

MIA2
Melanoma inhibitory activity 2
SEQ ID NO: 8712

MIA3
Melanoma inhibitory activity family, member 3
SEQ ID NOS: 8713-8717

MICU1
Mitochondrial calcium uptake 1
SEQ ID NOS: 8718-8727

MIER1
Mesoderm induction early response 1,
SEQ ID NOS: 8728-8736

transcriptional regulator

MINOS1-
MINOS1-NBL1 readthrough
SEQ ID NOS: 8737-8739

NBL1

MINPP1
Multiple inositol-polyphosphate phosphatase 1
SEQ ID NOS: 8740-8742

MLEC
Malectin
SEQ ID NOS: 8743-8746

MLN
Motilin
SEQ ID NOS: 8747-8749

MLXIP
MLX interacting protein
SEQ ID NOS: 8750-8755

MLXIPL
MLX interacting protein-like
SEQ ID NOS: 8756-8763

MMP1
Matrix metallopeptidase 1
SEQ ID NO: 8764

MMP10
Matrix metallopeptidase 10
SEQ ID NOS: 8765-8766

MMP11
Matrix metallopeptidase 11
SEQ ID NOS: 8767-8770

MMP12
Matrix metallopeptidase 12
SEQ ID NO: 8771

MMP13
Matrix metallopeptidase 13
SEQ ID NOS: 8772-8774

MMP14
Matrix metallopeptidase 14 (membrane-inserted)
SEQ ID NOS: 8775-8777

MMP17
Matrix metallopeptidase 17 (membrane-inserted)
SEQ ID NOS: 8778-8785

MMP19
Matrix metallopeptidase 19
SEQ ID NOS: 8786-8791

MMP2
Matrix metallopeptidase 2
SEQ ID NOS: 8792-8799

MMP20
Matrix metallopeptidase 20
SEQ ID NO: 8800

MMP21
Matrix metallopeptidase 21
SEQ ID NO: 8801

MMP25
Matrix metallopeptidase 25
SEQ ID NOS: 8802-8803

MMP26
Matrix metallopeptidase 26
SEQ ID NOS: 8804-8805

MMP27
Matrix metallopeptidase 27
SEQ ID NO: 8806

MMP28
Matrix metallopeptidase 28
SEQ ID NOS: 8807-8812

MMP3
Matrix metallopeptidase 3
SEQ ID NOS: 8813-8815

MMP7
Matrix metallopeptidase 7
SEQ ID NO: 8816

MMP8
Matrix metallopeptidase 8
SEQ ID NOS: 8817-8822

MMP9
Matrix metallopeptidase 9
SEQ ID NO: 8823

MMRN1
Multimerin 1
SEQ ID NOS: 8824-8826

MMRN2
Multimerin 2
SEQ ID NOS: 8827-8831

MOXD1
Monooxygenase, DBH-like 1
SEQ ID NOS: 8832-8834

C6orf25
Chromosome 6 open reading frame 25
SEQ ID NOS: 8835-8842

MPO
Myeloperoxidase
SEQ ID NOS: 8843-8844

MPPED1
Metallophosphoesterase domain containing 1
SEQ ID NOS: 8845-8848

MPZL1
Myelin protein zero-like 1
SEQ ID NOS: 8849-8853

MR1
Major histocompatibility complex, class I-related
SEQ ID NOS: 8854-8859

MRPL2
Mitochondrial ribosomal protein L2
SEQ ID NOS: 8860-8864

MRPL21
Mitochondrial ribosomal protein L21
SEQ ID NOS: 8865-8871

MRPL22
Mitochondrial ribosomal protein L22
SEQ ID NOS: 8872-8876

MRPL24
Mitochondrial ribosomal protein L24
SEQ ID NOS: 8877-8881

MRPL27
Mitochondrial ribosomal protein L27
SEQ ID NOS: 8882-8887

MRPL32
Mitochondrial ribosomal protein L32
SEQ ID NOS: 8888-8890

MRPL34
Mitochondrial ribosomal protein L34
SEQ ID NOS: 8891-8895

MRPL35
Mitochondrial ribosomal protein L35
SEQ ID NOS: 8896-8899

MRPL52
Mitochondrial ribosomal protein L52
SEQ ID NOS: 8900-8910

MRPL55
Mitochondrial ribosomal protein L55
SEQ ID NOS: 8911-8936

MRPS14
Mitochondrial ribosomal protein S14
SEQ ID NOS: 8937-8938

MRPS22
Mitochondrial ribosomal protein S22
SEQ ID NOS: 8939-8947

MRPS28
Mitochondrial ribosomal protein S28
SEQ ID NOS: 8948-8955

MS4A14
Membrane-spanning 4-domains, subfamily A,
SEQ ID NOS: 8956-8966

member 14

MS4A3
Membrane-spanning 4-domains, subfamily A,
SEQ ID NOS: 8967-8971

member 3 (hematopoietic cell-specific)

MSH3
MutS homolog 3
SEQ ID NO: 8972

MSH5
MutS homolog 5
SEQ ID NOS: 8973-8984

MSLN
Mesothelin
SEQ ID NOS: 8985-8992

MSMB
Microseminoprotein, beta-
SEQ ID NOS: 8993-8994

MSRA
Methionine sulfoxide reductase A
SEQ ID NOS: 8995-9002

MSRB2
Methionine sulfoxide reductase B2
SEQ ID NOS: 9003-9004

MSRB3
Methionine sulfoxide reductase B3
SEQ ID NOS: 9005-9018

MST1
Macrophage stimulating 1
SEQ ID NOS: 9019-9020

MSTN
Myostatin
SEQ ID NO: 9021

MT1G
Metallothionein 1G
SEQ ID NOS: 9022-9025

MTHFD2
Methylenetetrahydrofolate dehydrogenase (NADP +
SEQ ID NOS: 9026-9030

dependent) 2, methenyltetrahydrofolate

cyclohydrolase

MTMR14
Myotubularin related protein 14
SEQ ID NOS: 9031-9041

MTRNR2L11
MT-RNR2-like 11 (pseudogene)
SEQ ID NO: 9042

MTRR
5-methyltetrahydrofolate-homocysteine
SEQ ID NOS: 9043-9055

methyltransferase reductase

MTTP
Microsomal triglyceride transfer protein
SEQ ID NOS: 9056-9066

MTX2
Metaxin 2
SEQ ID NOS: 9067-9071

MUC1
Mucin 1, cell surface associated
SEQ ID NOS: 9072-9097

MUC13
Mucin 13, cell surface associated
SEQ ID NOS: 9098-9099

MUC20
Mucin 20, cell surface associated
SEQ ID NOS: 9100-9104

MUC3A
Mucin 3A, cell surface associated
SEQ ID NOS: 9105-9107

MUC5AC
Mucin 5AC, oligomeric mucus/gel-forming
SEQ ID NO: 9108

MUC5B
Mucin 5B, oligomeric mucus/gel-forming
SEQ ID NOS: 9109-9110

MUC6
Mucin 6, oligomeric mucus/gel-forming
SEQ ID NOS: 9111-9114

MUC7
Mucin 7, secreted
SEQ ID NOS: 9115-9118

MUCL1
Mucin-like 1
SEQ ID NOS: 9119-9121

MXRA5
Matrix-remodelling associated 5
SEQ ID NO: 9122

MXRA7
Matrix-remodelling associated 7
SEQ ID NOS: 9123-9129

MYDGF
Myeloid-derived growth factor
SEQ ID NOS: 9130-9132

MYL1
Myosin, light chain 1, alkali; skeletal, fast
SEQ ID NOS: 9133-9134

MYOC
Myocilin, trabecular meshwork inducible
SEQ ID NOS: 9135-9136

glucocorticoid response

MYRFL
Myelin regulatory factor-like
SEQ ID NOS: 9137-9141

MZB1
Marginal zone B and B1 cell-specific protein
SEQ ID NOS: 9142-9146

N4BP2L2
NEDD4 binding protein 2-like 2
SEQ ID NOS: 9147-9152

NAA38
N(alpha)-acetyltransferase 38, NatC auxiliary subunit
SEQ ID NOS: 9153-9158

NAAA
N-acylethanolamine acid amidase
SEQ ID NOS: 9159-9164

NAGA
N-acetylgalactosaminidase, alpha-
SEQ ID NOS: 9165-9167

NAGLU
N-acetylglucosaminidase, alpha
SEQ ID NOS: 9168-9172

NAGS
N-acetylglutamate synthase
SEQ ID NOS: 9173-9174

NAPSA
Napsin A aspartic peptidase
SEQ ID NOS: 9175-9177

CARKD
Carbohydrate kinase domain containing
SEQ ID NOS: 9178-9179

APOA1BP
Apolipoprotein A-I binding protein
SEQ ID NOS: 9180-9182

NBL1
Neuroblastoma 1, DAN family BMP antagonist
SEQ ID NOS: 9183-9196

NCAM1
Neural cell adhesion molecule 1
SEQ ID NOS: 9197-9216

NCAN
Neurocan
SEQ ID NOS: 9217-9218

NCBP2-AS2
NCBP2 antisense RNA 2 (head to head)
SEQ ID NO: 9219

NCSTN
Nicastrin
SEQ ID NOS: 9220-9229

NDNF
Neuron-derived neurotrophic factor
SEQ ID NOS: 9230-9232

NDP
Norrie disease (pseudoglioma)
SEQ ID NOS: 9233-9235

NDUFA10
NADH dehydrogenase (ubiquinone) 1 alpha
SEQ ID NOS: 9236-9245

subcomplex, 10, 42 kDa

NDUFB5
NADH dehydrogenase (ubiquinone) 1 beta
SEQ ID NOS: 9246-9254

subcomplex, 5, 16 kDa

NDUFS8
NADH dehydrogenase (ubiquinone) Fe—S protein 8,
SEQ ID NOS: 9255-9264

23 kDa (NADH-coenzyme Q reductase)

NDUFV1
NADH dehydrogenase (ubiquinone) flavoprotein 1,
SEQ ID NOS: 9265-9278

51 kDa

NECAB3
N-terminal EF-hand calcium binding protein 3
SEQ ID NOS: 9279-9288

PVRL1
Poliovirus receptor-related 1 (herpesvirus entry
SEQ ID NOS: 9289-9291

mediator C)

NELL1
Neural EGFL like 1
SEQ ID NOS: 9292-9295

NELL2
Neural EGFL like 2
SEQ ID NOS: 9296-9310

NENF
Neudesin neurotrophic factor
SEQ ID NO: 9311

NETO1
Neuropilin (NRP) and tolloid (TLL)-like 1
SEQ ID NOS: 9312-9316

NFASC
Neurofascin
SEQ ID NOS: 9317-9331

NFE2L1
Nuclear factor, erythroid 2-like 1
SEQ ID NOS: 9332-9350

NFE2L3
Nuclear factor, erythroid 2-like 3
SEQ ID NOS: 9351-9352

NGEF
Neuronal guanine nucleotide exchange factor
SEQ ID NOS: 9353-9358

NGF
Nerve growth factor (beta polypeptide)
SEQ ID NO: 9359

NGLY1
N-glycanase 1
SEQ ID NOS: 9360-9366

NGRN
Neugrin, neurite outgrowth associated
SEQ ID NOS: 9367-9368

NHLRC3
NHL repeat containing 3
SEQ ID NOS: 9369-9371

NID1
Nidogen 1
SEQ ID NOS: 9372-9373

NID2
Nidogen 2 (osteonidogen)
SEQ ID NOS: 9374-9376

NKG7
Natural killer cell granule protein 7
SEQ ID NOS: 9377-9381

NLGN3
Neuroligin 3
SEQ ID NOS: 9382-9386

NLGN4Y
Neuroligin 4, Y-linked
SEQ ID NOS: 9387-9393

NLRP5
NLR family, pyrin domain containing 5
SEQ ID NOS: 9394-9396

NMB
Neuromedin B
SEQ ID NOS: 9397-9398

NME1
NME/NM23 nucleoside diphosphate kinase 1
SEQ ID NOS: 9399-9405

NME1-
NME1-NME2 readthrough
SEQ ID NOS: 9406-9408

NME2

NME3
NME/NM23 nucleoside diphosphate kinase 3
SEQ ID NOS: 9409-9413

NMS
Neuromedin S
SEQ ID NO: 9414

NMU
Neuromedin U
SEQ ID NOS: 9415-9418

NOA1
Nitric oxide associated 1
SEQ ID NO: 9419

NODAL
Nodal growth differentiation factor
SEQ ID NOS: 9420-9421

NOG
Noggin
SEQ ID NO: 9422

NOMO3
NODAL modulator 3
SEQ ID NOS: 9423-9429

NOS1AP
Nitric oxide synthase 1 (neuronal) adaptor protein
SEQ ID NOS: 9430-9434

NOTCH3
Notch 3
SEQ ID NOS: 9435-9438

NOTUM
Notum pectinacetylesterase homolog (Drosophila)
SEQ ID NOS: 9439-9441

NOV
Nephroblastoma overexpressed
SEQ ID NO: 9442

NPB
Neuropeptide B
SEQ ID NOS: 9443-9444

NPC2
Niemann-Pick disease, type C2
SEQ ID NOS: 9445-9453

NPFF
Neuropeptide FF-amide peptide precursor
SEQ ID NO: 9454

NPFFR2
Neuropeptide FF receptor 2
SEQ ID NOS: 9455-9458

NPHS1
Nephrosis 1, congenital, Finnish type (nephrin)
SEQ ID NOS: 9459-9460

NPNT
Nephronectin
SEQ ID NOS: 9461-9471

NPPA
Natriuretic peptide A
SEQ ID NOS: 9472-9474

NPPB
Natriuretic peptide B
SEQ ID NO: 9475

NPPC
Natriuretic peptide C
SEQ ID NOS: 9476-9477

NPS
Neuropeptide S
SEQ ID NO: 9478

NPTX1
Neuronal pentraxin I
SEQ ID NO: 9479

NPTX2
Neuronal pentraxin II
SEQ ID NO: 9480

NPTXR
Neuronal pentraxin receptor
SEQ ID NOS: 9481-9482

NPVF
Neuropeptide VF precursor
SEQ ID NO: 9483

NPW
Neuropeptide W
SEQ ID NOS: 9484-9486

NPY
Neuropeptide Y
SEQ ID NOS: 9487-9489

NQO2
NAD(P)H dehydrogenase, quinone 2
SEQ ID NOS: 9490-9498

NRCAM
Neuronal cell adhesion molecule
SEQ ID NOS: 9499-9511

NRG1
Neuregulin 1
SEQ ID NOS: 9512-9529

NRN1L
Neuritin 1-like
SEQ ID NOS: 9530-9532

NRP1
Neuropilin 1
SEQ ID NOS: 9533-9546

NRP2
Neuropilin 2
SEQ ID NOS: 9547-9553

NRTN
Neurturin
SEQ ID NO: 9554

NRXN1
Neurexin 1
SEQ ID NOS: 9555-9585

NRXN2
Neurexin 2
SEQ ID NOS: 9586-9594

NT5C3A
5′-nucleotidase, cytosolic IIIA
SEQ ID NOS: 9595-9605

NT5DC3
5′-nucleotidase domain containing 3
SEQ ID NOS: 9606-9608

NT5E
5′-nucleotidase, ecto (CD73)
SEQ ID NOS: 9609-9613

NTF3
Neurotrophin 3
SEQ ID NOS: 9614-9615

NTF4
Neurotrophin 4
SEQ ID NOS: 9616-9617

NTM
Neurotrimin
SEQ ID NOS: 9618-9627

NTN1
Netrin 1
SEQ ID NOS: 9628-9629

NTN3
Netrin 3
SEQ ID NO: 9630

NTN4
Netrin 4
SEQ ID NOS: 9631-9635

NTN5
Netrin 5
SEQ ID NOS: 9636-9637

NTNG1
Netrin G1
SEQ ID NOS: 9638-9644

NTNG2
Netrin G2
SEQ ID NOS: 9645-9646

NTS
Neurotensin
SEQ ID NOS: 9647-9648

NUBPL
Nucleotide binding protein-like
SEQ ID NOS: 9649-9655

NUCB1
Nucleobindin 1
SEQ ID NOS: 9656-9662

NUCB2
Nucleobindin 2
SEQ ID NOS: 9663-9678

NUDT19
Nudix (nucleoside diphosphate linked moiety X)-type
SEQ ID NO: 9679

motif 19

NUDT9
Nudix (nucleoside diphosphate linked moiety X)-type
SEQ ID NOS: 9680-9684

motif 9

NUP155
Nucleoporin 155 kDa
SEQ ID NOS: 9685-9688

NUP214
Nucleoporin 214 kDa
SEQ ID NOS: 9689-9700

NUP85
Nucleoporin 85 kDa
SEQ ID NOS: 9701-9715

NXPE3
Neurexophilin and PC-esterase domain family,
SEQ ID NOS: 9716-9721

member 3

NXPE4
Neurexophilin and PC-esterase domain family,
SEQ ID NOS: 9722-9723

member 4

NXPH1
Neurexophilin 1
SEQ ID NOS: 9724-9727

NXPH2
Neurexophilin 2
SEQ ID NO: 9728

NXPH3
Neurexophilin 3
SEQ ID NOS: 9729-9730

NXPH4
Neurexophilin 4
SEQ ID NOS: 9731-9732

NYX
Nyctalopin
SEQ ID NOS: 9733-9734

OAF
Out at first homolog
SEQ ID NOS: 9735-9736

OBP2A
Odorant binding protein 2A
SEQ ID NOS: 9737-9743

OBP2B
Odorant binding protein 2B
SEQ ID NOS: 9744-9747

OC90
Otoconin 90
SEQ ID NO: 9748

OCLN
Occludin
SEQ ID NOS: 9749-9751

ODAM
Odontogenic, ameloblast asssociated
SEQ ID NOS: 9752-9755

C4orf26
Chromosome 4 open reading frame 26
SEQ ID NOS: 9756-9759

OGG1
8-oxoguanine DNA glycosylase
SEQ ID NOS: 9760-9773

OGN
Osteoglycin
SEQ ID NOS: 9774-9776

OIT3
Oncoprotein induced transcript 3
SEQ ID NOS: 9777-9778

OLFM1
Olfactomedin 1
SEQ ID NOS: 9779-9789

OLFM2
Olfactomedin 2
SEQ ID NOS: 9790-9793

OLFM3
Olfactomedin 3
SEQ ID NOS: 9794-9796

OLFM4
Olfactomedin 4
SEQ ID NO: 9797

OLFML1
Olfactomedin-like 1
SEQ ID NOS: 9798-9801

OLFML2A
Olfactomedin-like 2A
SEQ ID NOS: 9802-9804

OLFML2B
Olfactomedin-like 2B
SEQ ID NOS: 9805-9809

OLFML3
Olfactomedin-like 3
SEQ ID NOS: 9810-9812

OMD
Osteomodulin
SEQ ID NO: 9813

OMG
Oligodendrocyte myelin glycoprotein
SEQ ID NO: 9814

OOSP2
Oocyte secreted protein 2
SEQ ID NOS: 9815-9816

OPCML
Opioid binding protein/cell adhesion molecule-like
SEQ ID NOS: 9817-9821

PROL1
Proline rich, lacrimal 1
SEQ ID NO: 9822

OPTC
Opticin
SEQ ID NOS: 9823-9824

ORAI1
ORAI calcium release-activated calcium modulator 1
SEQ ID NO: 9825

ORM1
Orosomucoid 1
SEQ ID NO: 9826

ORM2
Orosomucoid 2
SEQ ID NO: 9827

ORMDL2
ORMDL sphingolipid biosynthesis regulator 2
SEQ ID NOS: 9828-9831

OS9
Osteosarcoma amplified 9, endoplasmic reticulum
SEQ ID NOS: 9832-9846

lectin

OSCAR
Osteoclast associated, immunoglobulin-like receptor
SEQ ID NOS: 9847-9857

OSM
Oncostatin M
SEQ ID NOS: 9858-9860

OSMR
Oncostatin M receptor
SEQ ID NOS: 9861-9865

OSTN
Osteocrin
SEQ ID NOS: 9866-9867

OTOA
Otoancorin
SEQ ID NOS: 9868-9873

OTOG
Otogelin
SEQ ID NOS: 9874-9876

OTOGL
Otogelin-like
SEQ ID NOS: 9877-9883

OTOL1
Otolin 1
SEQ ID NO: 9884

OTOR
Otoraplin
SEQ ID NO: 9885

OTOS
Otospiralin
SEQ ID NOS: 9886-9887

OVCH1
Ovochymase 1
SEQ ID NOS: 9888-9890

OVCH2
Ovochymase 2 (gene/pseudogene)
SEQ ID NOS: 9891-9892

OVGP1
Oviductal glycoprotein 1, 120 kDa
SEQ ID NO: 9893

OXCT1
3-oxoacid CoA transferase 1
SEQ ID NOS: 9894-9897

OXCT2
3-oxoacid CoA transferase 2
SEQ ID NO: 9898

OXNAD1
Oxidoreductase NAD-binding domain containing 1
SEQ ID NOS: 9899-9905

OXT
Oxytocin/neurophysin I prepropeptide
SEQ ID NO: 9906

P3H1
Prolyl 3-hydroxylase 1
SEQ ID NOS: 9907-9911

P3H2
Prolyl 3-hydroxylase 2
SEQ ID NOS: 9912-9915

P3H3
Prolyl 3-hydroxylase 3
SEQ ID NO: 9916

P3H4
Prolyl 3-hydroxylase family member 4 (non-
SEQ ID NOS: 9917-9921

enzymatic)

P4HA1
Prolyl 4-hydroxylase, alpha polypeptide I
SEQ ID NOS: 9922-9926

P4HA2
Prolyl 4-hydroxylase, alpha polypeptide II
SEQ ID NOS: 9927-9941

P4HA3
Prolyl 4-hydroxylase, alpha polypeptide III
SEQ ID NOS: 9942-9946

P4HB
Prolyl 4-hydroxylase, beta polypeptide
SEQ ID NOS: 9947-9958

PAEP
Progestagen-associated endometrial protein
SEQ ID NOS: 9959-9967

PAM
Peptidylglycine alpha-amidating monooxygenase
SEQ ID NOS: 9968-9981

PAMR1
Peptidase domain containing associated with muscle
SEQ ID NOS: 9982-9988

regeneration 1

PAPLN
Papilin, proteoglycan-like sulfated glycoprotein
SEQ ID NOS: 9989-9996

PAPPA
Pregnancy-associated plasma protein A,
SEQ ID NO: 9997

pappalysin 1

PAPPA2
Pappalysin 2
SEQ ID NOS: 9998-9999

PARP15
Poly (ADP-ribose) polymerase family, member 15
SEQ ID NOS: 10000-10003

PARVB
Parvin, beta
SEQ ID NOS: 10004-10008

PATE1
Prostate and testis expressed 1
SEQ ID NOS: 10009-10010

PATE2
Prostate and testis expressed 2
SEQ ID NOS: 10011-10012

PATE3
Prostate and testis expressed 3
SEQ ID NO: 10013

PATE4
Prostate and testis expressed 4
SEQ ID NOS: 10014-10015

PATL2
Protein associated with topoisomerase II homolog 2
SEQ ID NOS: 10016-10021

(yeast)

PAX2
Paired box 2
SEQ ID NOS: 10022-10027

PAX4
Paired box 4
SEQ ID NOS: 10028-10034

PCCB
Propionyl CoA carboxylase, beta polypeptide
SEQ ID NOS: 10035-10049

PCDH1
Protocadherin 1
SEQ ID NOS: 10050-10055

PCDH12
Protocadherin 12
SEQ ID NOS: 10056-10057

PCDH15
Protocadherin-related 15
SEQ ID NOS: 10058-10091

PCDHA1
Protocadherin alpha 1
SEQ ID NOS: 10092-10094

PCDHA10
Protocadherin alpha 10
SEQ ID NOS: 10095-10097

PCDHA11
Protocadherin alpha 11
SEQ ID NOS: 10098-10100

PCDHA6
Protocadherin alpha 6
SEQ ID NOS: 10101-10103

PCDHB12
Protocadherin beta 12
SEQ ID NOS: 10104-10106

PCDHGA11
Protocadherin gamma subfamily A, 11
SEQ ID NOS: 10107-10109

PCF11
PCF11 cleavage and polyadenylation factor subunit
SEQ ID NOS: 10110-10114

PCOLCE
Procollagen C-endopeptidase enhancer
SEQ ID NO: 10115

PCOLCE2
Procollagen C-endopeptidase enhancer 2
SEQ ID NOS: 10116-10119

PCSK1
Proprotein convertase subtilisin/kexin type 1
SEQ ID NOS: 10120-10122

PCSK1N
Proprotein convertase subtilisin/kexin type 1
SEQ ID NO: 10123

inhibitor

PCSK2
Proprotein convertase subtilisin/kexin type 2
SEQ ID NOS: 10124-10126

PCSK4
Proprotein convertase subtilisin/kexin type 4
SEQ ID NOS: 10127-10129

PCSK5
Proprotein convertase subtilisin/kexin type 5
SEQ ID NOS: 10130-10134

PCSK9
Proprotein convertase subtilisin/kexin type 9
SEQ ID NO: 10135

PCYOX1
Prenylcysteine oxidase 1
SEQ ID NOS: 10136-10140

PCYOX1L
Prenylcysteine oxidase 1 like
SEQ ID NOS: 10141-10145

PDE11A
Phosphodiesterase 11A
SEQ ID NOS: 10146-10151

PDE2A
Phosphodiesterase 2A, cGMP-stimulated
SEQ ID NOS: 10152-10173

PDE7A
Phosphodiesterase 7A
SEQ ID NOS: 10174-10177

PDF
Peptide deformylase (mitochondrial)
SEQ ID NO: 10178

PDGFA
Platelet-derived growth factor alpha polypeptide
SEQ ID NOS: 10179-10182

PDGFB
Platelet-derived growth factor beta polypeptide
SEQ ID NOS: 10183-10186

PDGFC
Platelet derived growth factor C
SEQ ID NOS: 10187-10190

PDGFD
Platelet derived growth factor D
SEQ ID NOS: 10191-10193

PDGFRA
Platelet-derived growth factor receptor, alpha
SEQ ID NOS: 10194-10200

polypeptide

PDGFRB
Platelet-derived growth factor receptor, beta
SEQ ID NOS: 10201-10204

polypeptide

PDGFRL
Platelet-derived growth factor receptor-like
SEQ ID NOS: 10205-10206

PDHA1
Pyruvate dehydrogenase (lipoamide) alpha 1
SEQ ID NOS: 10207-10215

PDIA2
Protein disulfide isomerase family A, member 2
SEQ ID NOS: 10216-10219

PDIA3
Protein disulfide isomerase family A, member 3
SEQ ID NOS: 10220-10223

PDIA4
Protein disulfide isomerase family A, member 4
SEQ ID NOS: 10224-10225

PDIA5
Protein disulfide isomerase family A, member 5
SEQ ID NOS: 10226-10229

PDIA6
Protein disulfide isomerase family A, member 6
SEQ ID NOS: 10230-10236

PDILT
Protein disulfide isomerase-like, testis expressed
SEQ ID NOS: 10237-10238

PDYN
Prodynorphin
SEQ ID NOS: 10239-10241

PDZD8
PDZ domain containing 8
SEQ ID NO: 10242

PDZRN4
PDZ domain containing ring finger 4
SEQ ID NOS: 10243-10245

PEAR1
Platelet endothelial aggregation receptor 1
SEQ ID NOS: 10246-10249

PEBP4
Phosphatidylethanolamine-binding protein 4
SEQ ID NOS: 10250-10251

PECAM1
Platelet/endothelial cell adhesion molecule 1
SEQ ID NOS: 10252-10255

PENK
Proenkephalin
SEQ ID NOS: 10256-10261

PET117
PET117 homolog
SEQ ID NO: 10262

PF4
Platelet factor 4
SEQ ID NO: 10263

PF4V1
Platelet factor 4 variant 1
SEQ ID NO: 10264

PFKP
Phosphofructokinase, platelet
SEQ ID NOS: 10265-10273

PFN1
Profilin 1
SEQ ID NOS: 10274-10276

PGA3
Pepsinogen 3, group I (pepsinogen A)
SEQ ID NOS: 10277-10280

PGA4
Pepsinogen 4, group I (pepsinogen A)
SEQ ID NOS: 10281-10283

PGA5
Pepsinogen 5, group I (pepsinogen A)
SEQ ID NOS: 10284-10286

PGAM5
PGAM family member 5, serine/threonine protein
SEQ ID NOS: 10287-10290

phosphatase, mitochondrial

PGAP3
Post-GPI attachment to proteins 3
SEQ ID NOS: 10291-10298

PGC
Progastricsin (pepsinogen C)
SEQ ID NOS: 10299-10302

PGF
Placental growth factor
SEQ ID NOS: 10303-10306

PGLYRP1
Peptidoglycan recognition protein 1
SEQ ID NO: 10307

PGLYRP2
Peptidoglycan recognition protein 2
SEQ ID NOS: 10308-10311

PGLYRP3
Peptidoglycan recognition protein 3
SEQ ID NO: 10312

PGLYRP4
Peptidoglycan recognition protein 4
SEQ ID NOS: 10313-10314

PHACTR1
Phosphatase and actin regulator 1
SEQ ID NOS: 10315-10321

PHB
Prohibitin
SEQ ID NOS: 10322-10330

PI15
Peptidase inhibitor 15
SEQ ID NOS: 10331-10332

PI3
Peptidase inhibitor 3, skin-derived
SEQ ID NO: 10333

PIANP
PILR alpha associated neural protein
SEQ ID NOS: 10334-10339

PIGK
Phosphatidylinositol glycan anchor biosynthesis,
SEQ ID NOS: 10340-10343

class K

PIGL
Phosphatidylinositol glycan anchor biosynthesis,
SEQ ID NOS: 10344-10351

class L

PIGT
Phosphatidylinositol glycan anchor biosynthesis,
SEQ ID NOS: 10352-10406

class T

PIGZ
Phosphatidylinositol glycan anchor biosynthesis,
SEQ ID NOS: 10407-10409

class Z

PIK3AP1
Phosphoinositide-3-kinase adaptor protein 1
SEQ ID NOS: 10410-10412

PIK3IP1
Phosphoinositide-3-kinase interacting protein 1
SEQ ID NOS: 10413-10416

PILRA
Paired immunoglobin-like type 2 receptor alpha
SEQ ID NOS: 10417-10421

PILRB
Paired immunoglobin-like type 2 receptor beta
SEQ ID NOS: 10422-10433

PINLYP
Phospholipase A2 inhibitor and LY6/PLAUR domain
SEQ ID NOS: 10434-10438

containing

PIP
Prolactin-induced protein
SEQ ID NO: 10439

PIWIL4
Piwi-like RNA-mediated gene silencing 4
SEQ ID NOS: 10440-10444

PKDCC
Protein kinase domain containing, cytoplasmic
SEQ ID NOS: 10445-10446

PKHD1
Polycystic kidney and hepatic disease 1 (autosomal
SEQ ID NOS: 10447-10448

recessive)

PLA1A
Phospholipase A1 member A
SEQ ID NOS: 10449-10453

PLA2G10
Phospholipase A2, group X
SEQ ID NOS: 10454-10455

PLA2G12A
Phospholipase A2, group XIIA
SEQ ID NOS: 10456-10458

PLA2G12B
Phospholipase A2, group XIIB
SEQ ID NO: 10459

PLA2G15
Phospholipase A2, group XV
SEQ ID NOS: 10460-10467

PLA2G1B
Phospholipase A2, group IB (pancreas)
SEQ ID NOS: 10468-10470

PLA2G2A
Phospholipase A2, group IIA (platelets, synovial
SEQ ID NOS: 10471-10472

fluid)

PLA2G2C
Phospholipase A2, group IIC
SEQ ID NOS: 10473-10474

PLA2G2D
Phospholipase A2, group IID
SEQ ID NOS: 10475-10476

PLA2G2E
Phospholipase A2, group IIE
SEQ ID NO: 10477

PLA2G3
Phospholipase A2, group III
SEQ ID NO: 10478

PLA2G5
Phospholipase A2, group V
SEQ ID NO: 10479

PLA2G7
Phospholipase A2, group VII (platelet-activating
SEQ ID NOS: 10480-10481

factor acetylhydrolase, plasma)

PLA2R1
Phospholipase A2 receptor 1, 180 kDa
SEQ ID NOS: 10482-10483

PLAC1
Placenta-specific 1
SEQ ID NO: 10484

PLAC9
Placenta-specific 9
SEQ ID NOS: 10485-10487

PLAT
Plasminogen activator, tissue
SEQ ID NOS: 10488-10496

PLAU
Plasminogen activator, urokinase
SEQ ID NOS: 10497-10499

PLAUR
Plasminogen activator, urokinase receptor
SEQ ID NOS: 10500-10511

PLBD1
Phospholipase B domain containing 1
SEQ ID NOS: 10512-10514

PLBD2
Phospholipase B domain containing 2
SEQ ID NOS: 10515-10517

PLG
Plasminogen
SEQ ID NOS: 10518-10520

PLGLB1
Plasminogen-like B1
SEQ ID NOS: 10521-10524

PLGLB2
Plasminogen-like B2
SEQ ID NOS: 10525-10526

PLOD1
Procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1
SEQ ID NOS: 10527-10529

PLOD2
Procollagen-lysine, 2-oxoglutarate 5-dioxygenase 2
SEQ ID NOS: 10530-10535

PLOD3
Procollagen-lysine, 2-oxoglutarate 5-dioxygenase 3
SEQ ID NOS: 10536-10542

PLTP
Phospholipid transfer protein
SEQ ID NOS: 10543-10547

PLXNA4
Plexin A4
SEQ ID NOS: 10548-10551

PLXNB2
Plexin B2
SEQ ID NOS: 10552-10560

PM20D1
Peptidase M20 domain containing 1
SEQ ID NO: 10561

PMCH
Pro-melanin-concentrating hormone
SEQ ID NO: 10562

PMEL
Premelanosome protein
SEQ ID NOS: 10563-10574

PMEPA1
Prostate transmembrane protein, androgen
SEQ ID NOS: 10575-10581

induced 1

PNLIP
Pancreatic lipase
SEQ ID NO: 10582

PNLIPRP1
Pancreatic lipase-related protein 1
SEQ ID NOS: 10583-10591

PNLIPRP3
Pancreatic lipase-related protein 3
SEQ ID NO: 10592

PNOC
Prepronociceptin
SEQ ID NOS: 10593-10595

PNP
Purine nucleoside phosphorylase
SEQ ID NOS: 10596-10599

PNPLA4
Patatin-like phospholipase domain containing 4
SEQ ID NOS: 10600-10603

PODNL1
Podocan-like 1
SEQ ID NOS: 10604-10615

POFUT1
Protein O-fucosyltransferase 1
SEQ ID NOS: 10616-10617

POFUT2
Protein O-fucosyltransferase 2
SEQ ID NOS: 10618-10623

POGLUT1
Protein O-glucosyltransferase 1
SEQ ID NOS: 10624-10628

POLL
Polymerase (DNA directed), lambda
SEQ ID NOS: 10629-10641

POMC
Proopiomelanocortin
SEQ ID NOS: 10642-10646

POMGNT2
Protein O-linked mannose N-
SEQ ID NOS: 10647-10648

acetylglucosaminyltransferase 2 (beta 1,4-)

PON1
Paraoxonase 1
SEQ ID NOS: 10649-10650

PON2
Paraoxonase 2
SEQ ID NOS: 10651-10663

PON3
Paraoxonase 3
SEQ ID NOS: 10664-10669

POSTN
Periostin, osteoblast specific factor
SEQ ID NOS: 10670-10675

PPBP
Pro-platelet basic protein (chemokine (C-X-C motif)
SEQ ID NO: 10676

ligand 7)

PPIB
Peptidylprolyl isomerase B (cyclophilin B)
SEQ ID NO: 10677

PPIC
Peptidylprolyl isomerase C (cyclophilin C)
SEQ ID NO: 10678

PPOX
Protoporphyrinogen oxidase
SEQ ID NOS: 10679-10689

PPP1CA
Protein phosphatase 1, catalytic subunit, alpha
SEQ ID NOS: 10690-10695

isozyme

PPT1
Palmitoyl-protein thioesterase 1
SEQ ID NOS: 10696-10712

PPT2
Palmitoyl-protein thioesterase 2
SEQ ID NOS: 10713-10720

PPY
Pancreatic polypeptide
SEQ ID NOS: 10721-10725

PRAC2
Prostate cancer susceptibility candidate 2
SEQ ID NOS: 10726-10727

PRADC1
Protease-associated domain containing 1
SEQ ID NO: 10728

PRAP1
Proline-rich acidic protein 1
SEQ ID NOS: 10729-10730

PRB1
Proline-rich protein BstNI subfamily 1
SEQ ID NOS: 10731-10734

PRB2
Proline-rich protein BstNI subfamily 2
SEQ ID NOS: 10735-10736

PRB3
Proline-rich protein BstNI subfamily 3
SEQ ID NOS: 10737-10738

PRB4
Proline-rich protein BstNI subfamily 4
SEQ ID NOS: 10739-10742

PRCD
Progressive rod-cone degeneration
SEQ ID NOS: 10743-10744

PRCP
Prolylcarboxypeptidase (angiotensinase C)
SEQ ID NOS: 10745-10756

PRDM12
PR domain containing 12
SEQ ID NO: 10757

PRDX4
Peroxiredoxin 4
SEQ ID NOS: 10758-10761

PRELP
Proline/arginine-rich end leucine-rich repeat protein
SEQ ID NO: 10762

PRF1
Perforin 1 (pore forming protein)
SEQ ID NOS: 10763-10765

PRG2
Proteoglycan 2, bone marrow (natural killer cell
SEQ ID NOS: 10766-10768

activator, eosinophil granule major basic protein)

PRG3
Proteoglycan 3
SEQ ID NO: 10769

PRG4
Proteoglycan 4
SEQ ID NOS: 10770-10775

PRH1
Proline-rich protein Haelll subfamily 1
SEQ ID NOS: 10776-10778

PRH2
Proline-rich protein Haelll subfamily 2
SEQ ID NOS: 10779-10780

PRKAG1
Protein kinase, AMP-activated, gamma 1 non-
SEQ ID NOS: 10781-10795

catalytic subunit

PRKCSH
Protein kinase C substrate 80K-H
SEQ ID NOS: 10796-10805

PRKD1
Protein kinase D1
SEQ ID NOS: 10806-10811

PRL
Prolactin
SEQ ID NOS: 10812-10814

PRLH
Prolactin releasing hormone
SEQ ID NO: 10815

PRLR
Prolactin receptor
SEQ ID NOS: 10816-10834

PRNP
Prion protein
SEQ ID NOS: 10835-10838

PRNT
Prion protein (testis specific)
SEQ ID NO: 10839

PROC
Protein C (inactivator of coagulation factors Va and
SEQ ID NOS: 10840-10847

VIIIa)

PROK1
Prokineticin 1
SEQ ID NO: 10848

PROK2
Prokineticin 2
SEQ ID NOS: 10849-10850

PROM1
Prominin 1
SEQ ID NOS: 10851-10862

PROS1
Protein S (alpha)
SEQ ID NOS: 10863-10866

PROZ
Protein Z, vitamin K-dependent plasma glycoprotein
SEQ ID NOS: 10867-10868

PRR27
Proline rich 27
SEQ ID NOS: 10869-10872

PRR4
Proline rich 4 (lacrimal)
SEQ ID NOS: 10873-10875

PRRG2
Proline rich Gla (G-carboxyglutamic acid) 2
SEQ ID NOS: 10876-10878

PRRT3
Proline-rich transmembrane protein 3
SEQ ID NOS: 10879-10881

PRRT4
Proline-rich transmembrane protein 4
SEQ ID NOS: 10882-10888

PRSS1
Protease, serine, 1 (trypsin 1)
SEQ ID NOS: 10889-10892

PRSS12
Protease, serine, 12 (neurotrypsin, motopsin)
SEQ ID NO: 10893

PRSS16
Protease, serine, 16 (thymus)
SEQ ID NOS: 10894-10901

PRSS2
Protease, serine, 2 (trypsin 2)
SEQ ID NOS: 10902-10905

PRSS21
Protease, serine, 21 (testisin)
SEQ ID NOS: 10906-10911

PRSS22
Protease, serine, 22
SEQ ID NOS: 10912-10914

PRSS23
Protease, serine, 23
SEQ ID NOS: 10915-10918

PRSS27
Protease, serine 27
SEQ ID NOS: 10919-10921

PRSS3
Protease, serine, 3
SEQ ID NOS: 10922-10926

PRSS33
Protease, serine, 33
SEQ ID NOS: 10927-10930

PRSS35
Protease, serine, 35
SEQ ID NO: 10931

PRSS36
Protease, serine, 36
SEQ ID NOS: 10932-10935

PRSS37
Protease, serine, 37
SEQ ID NOS: 10936-10939

PRSS38
Protease, serine, 38
SEQ ID NO: 10940

PRSS42
Protease, serine, 42
SEQ ID NOS: 10941-10942

PRSS48
Protease, serine, 48
SEQ ID NOS: 10943-10944

PRSS50
Protease, serine, 50
SEQ ID NO: 10945

PRSS53
Protease, serine, 53
SEQ ID NO: 10946

PRSS54
Protease, serine, 54
SEQ ID NOS: 10947-10951

PRSS55
Protease, serine, 55
SEQ ID NOS: 10952-10954

PRSS56
Protease, serine, 56
SEQ ID NOS: 10955-10956

PRSS57
Protease, serine, 57
SEQ ID NOS: 10957-10958

PRSS58
Protease, serine, 58
SEQ ID NOS: 10959-10960

PRSS8
Protease, serine, 8
SEQ ID NOS: 10961-10964

PRTG
Protogenin
SEQ ID NOS: 10965-10968

PRTN3
Proteinase 3
SEQ ID NOS: 10969-10970

PSAP
Prosaposin
SEQ ID NOS: 10971-10974

PSAPL1
Prosaposin-like 1 (gene/pseudogene)
SEQ ID NO: 10975

PSG1
Pregnancy specific beta-1-glycoprotein 1
SEQ ID NOS: 10976-10983

PSG11
Pregnancy specific beta-1-glycoprotein 11
SEQ ID NOS: 10984-10988

PSG2
Pregnancy specific beta-1-glycoprotein 2
SEQ ID NOS: 10989-10990

PSG3
Pregnancy specific beta-1-glycoprotein 3
SEQ ID NOS: 10991-10994

PSG4
Pregnancy specific beta-1-glycoprotein 4
SEQ ID NOS: 10995-11006

PSG5
Pregnancy specific beta-1-glycoprotein 5
SEQ ID NOS: 11007-11012

PSG6
Pregnancy specific beta-1-glycoprotein 6
SEQ ID NOS: 11013-11018

PSG7
Pregnancy specific beta-1-glycoprotein 7
SEQ ID NOS: 11019-11021

(gene/pseudogene)

PSG8
Pregnancy specific beta-1-glycoprotein 8
SEQ ID NOS: 11022-11026

PSG9
Pregnancy specific beta-1-glycoprotein 9
SEQ ID NOS: 11027-11034

PSMD1
Proteasome 26S subunit, non-ATPase 1
SEQ ID NOS: 11035-11042

PSORS1C2
Psoriasis susceptibility 1 candidate 2
SEQ ID NO: 11043

PSPN
Persephin
SEQ ID NOS: 11044-11045

PTGDS
Prostaglandin D2 synthase 21 kDa (brain)
SEQ ID NOS: 11046-11050

PTGIR
Prostaglandin I2 (prostacyclin) receptor (IP)
SEQ ID NOS: 11051-11055

PTGS1
Prostaglandin-endoperoxide synthase 1
SEQ ID NOS: 11056-11064

(prostaglandin G/H synthase and cyclooxygenase)

PTGS2
Prostaglandin-endoperoxide synthase 2
SEQ ID NOS: 11065-11066

(prostaglandin G/H synthase and cyclooxygenase)

PTH
Parathyroid hormone
SEQ ID NOS: 11067-11068

PTH2
Parathyroid hormone 2
SEQ ID NO: 11069

PTHLH
Parathyroid hormone-like hormone
SEQ ID NOS: 11070-11078

PTK7
Protein tyrosine kinase 7 (inactive)
SEQ ID NOS: 11079-11094

PTN
Pleiotrophin
SEQ ID NOS: 11095-11096

PTPRA
Protein tyrosine phosphatase, receptor type, A
SEQ ID NOS: 11097-11104

PTPRB
Protein tyrosine phosphatase, receptor type, B
SEQ ID NOS: 11105-11112

PTPRC
Protein tyrosine phosphatase, receptor type, C
SEQ ID NOS: 11113-11123

PTPRCAP
Protein tyrosine phosphatase, receptor type, C-
SEQ ID NO: 11124

associated protein

PTPRD
Protein tyrosine phosphatase, receptor type, D
SEQ ID NOS: 11125-11136

PTPRF
Protein tyrosine phosphatase, receptor type, F
SEQ ID NOS: 11137-11144

PTPRJ
Protein tyrosine phosphatase, receptor type, J
SEQ ID NOS: 11145-11150

PTPRO
Protein tyrosine phosphatase, receptor type, O
SEQ ID NOS: 11151-11159

PTPRS
Protein tyrosine phosphatase, receptor type, S
SEQ ID NOS: 11160-11167

PTTG1IP
Pituitary tumor-transforming 1 interacting protein
SEQ ID NOS: 11168-11171

PTX3
Pentraxin 3, long
SEQ ID NO: 11172

PTX4
Pentraxin 4, long
SEQ ID NOS: 11173-11175

PVR
Poliovirus receptor
SEQ ID NOS: 11176-11181

PXDN
Peroxidasin
SEQ ID NOS: 11182-11186

PXDNL
Peroxidasin-like
SEQ ID NOS: 11187-11189

PXYLP1
2-phosphoxylose phosphatase 1
SEQ ID NOS: 11190-11202

PYY
Peptide YY
SEQ ID NOS: 11203-11204

PZP
Pregnancy-zone protein
SEQ ID NOS: 11205-11206

QPCT
Glutaminyl-peptide cyclotransferase
SEQ ID NOS: 11207-11209

QPRT
Quinolinate phosphoribosyltransferase
SEQ ID NOS: 11210-11211

QRFP
Pyroglutamylated RFamide peptide
SEQ ID NOS: 11212-11213

QSOX1
Quiescin Q6 sulfhydryl oxidase 1
SEQ ID NOS: 11214-11217

R3HDML
R3H domain containing-like
SEQ ID NO: 11218

RAB26
RAB26, member RAS oncogene family
SEQ ID NOS: 11219-11222

RAB36
RAB36, member RAS oncogene family
SEQ ID NOS: 11223-11225

RAB9B
RAB9B, member RAS oncogene family
SEQ ID NO: 11226

RAET1E
Retinoic acid early transcript 1E
SEQ ID NOS: 11227-11232

RAET1G
Retinoic acid early transcript 1G
SEQ ID NOS: 11233-11235

RAMP2
Receptor (G protein-coupled) activity modifying
SEQ ID NOS: 11236-11240

protein 2

RAPGEF5
Rap guanine nucleotide exchange factor (GEF) 5
SEQ ID NOS: 11241-11247

RARRES1
Retinoic acid receptor responder (tazarotene
SEQ ID NOS: 11248-11249

induced) 1

RARRES2
Retinoic acid receptor responder (tazarotene
SEQ ID NOS: 11250-11253

induced) 2

RASA2
RAS p21 protein activator 2
SEQ ID NOS: 11254-11256

RBM3
RNA binding motif (RNP1, RRM) protein 3
SEQ ID NOS: 11257-11259

RBP3
Retinol binding protein 3, interstitial
SEQ ID NO: 11260

RBP4
Retinol binding protein 4, plasma
SEQ ID NOS: 11261-11264

RCN1
Reticulocalbin 1, EF-hand calcium binding domain
SEQ ID NOS: 11265-11268

RCN2
Reticulocalbin 2, EF-hand calcium binding domain
SEQ ID NOS: 11269-11272

RCN3
Reticulocalbin 3, EF-hand calcium binding domain
SEQ ID NOS: 11273-11276

RCOR1
REST corepressor 1
SEQ ID NOS: 11277-11278

RDH11
Retinol dehydrogenase 11 (all-trans/9-cis/11-cis)
SEQ ID NOS: 11279-11286

RDH12
Retinol dehydrogenase 12 (all-trans/9-cis/11-cis)
SEQ ID NOS: 11287-11288

RDH13
Retinol dehydrogenase 13 (all-trans/9-cis)
SEQ ID NOS: 11289-11297

RDH5
Retinol dehydrogenase 5 (11-cis/9-cis)
SEQ ID NOS: 11298-11302

RDH8
Retinol dehydrogenase 8 (all-trans)
SEQ ID NOS: 11303-11304

REG1A
Regenerating islet-derived 1 alpha
SEQ ID NO: 11305

REG1B
Regenerating islet-derived 1 beta
SEQ ID NOS: 11306-11307

REG3A
Regenerating islet-derived 3 alpha
SEQ ID NOS: 11308-11310

REG3G
Regenerating islet-derived 3 gamma
SEQ ID NOS: 11311-11313

REG4
Regenerating islet-derived family, member 4
SEQ ID NOS: 11314-11317

RELN
Reelin
SEQ ID NOS: 11318-11321

RELT
RELT tumor necrosis factor receptor
SEQ ID NOS: 11322-11325

REN
Renin
SEQ ID NOS: 11326-11327

REPIN1
Replication initiator 1
SEQ ID NOS: 11328-11341

REPS2
RALBP1 associated Eps domain containing 2
SEQ ID NOS: 11342-11343

RET
Ret proto-oncogene
SEQ ID NOS: 11344-11349

RETN
Resistin
SEQ ID NOS: 11350-11352

RETNLB
Resistin like beta
SEQ ID NO: 11353

RETSAT
Retinol saturase (all-trans-retinol 13,14-reductase)
SEQ ID NOS: 11354-11358

RFNG
RFNG O-fucosylpeptide 3-beta-N-
SEQ ID NOS: 11359-11361

acetylglucosaminyltransferase

RGCC
Regulator of cell cycle
SEQ ID NO: 11362

RGL4
Ral guanine nucleotide dissociation stimulator-like 4
SEQ ID NOS: 11363-11369

RGMA
Repulsive guidance molecule family member a
SEQ ID NOS: 11370-11379

RGMB
Repulsive guidance molecule family member b
SEQ ID NOS: 11380-11381

RHOQ
Ras homolog family member Q
SEQ ID NOS: 11382-11386

RIC3
RIC3 acetylcholine receptor chaperone
SEQ ID NOS: 11387-11394

HRSP12
Heat-responsive protein 12
SEQ ID NOS: 11395-11398

RIMS1
Regulating synaptic membrane exocytosis 1
SEQ ID NOS: 11399-11414

RIPPLY1
Ripply transcriptional repressor 1
SEQ ID NOS: 11415-11416

RLN1
Relaxin 1
SEQ ID NO: 11417

RLN2
Relaxin 2
SEQ ID NOS: 11418-11419

RLN3
Relaxin 3
SEQ ID NOS: 11420-11421

RMDN1
Regulator of microtubule dynamics 1
SEQ ID NOS: 11422-11435

RNASE1
Ribonuclease, RNase A family, 1 (pancreatic)
SEQ ID NOS: 11436-11440

RNASE10
Ribonuclease, RNase A family, 10 (non-active)
SEQ ID NOS: 11441-11442

RNASE11
Ribonuclease, RNase A family, 11 (non-active)
SEQ ID NOS: 11443-11453

RNASE12
Ribonuclease, RNase A family, 12 (non-active)
SEQ ID NO: 11454

RNASE13
Ribonuclease, RNase A family, 13 (non-active)
SEQ ID NO: 11455

RNASE2
Ribonuclease, RNase A family, 2 (liver, eosinophil-
SEQ ID NO: 11456

derived neurotoxin)

RNASE3
Ribonuclease, RNase A family, 3
SEQ ID NO: 11457

RNASE4
Ribonuclease, RNase A family, 4
SEQ ID NOS: 11458-11460

RNASE6
Ribonuclease, RNase A family, k6
SEQ ID NO: 11461

RNASE7
Ribonuclease, RNase A family, 7
SEQ ID NOS: 11462-11463

RNASE8
Ribonuclease, RNase A family, 8
SEQ ID NO: 11464

RNASE9
Ribonuclease, RNase A family, 9 (non-active)
SEQ ID NOS: 11465-11475

RNASEH1
Ribonuclease H1
SEQ ID NOS: 11476-11478

RNASET2
Ribonuclease T2
SEQ ID NOS: 11479-11486

RNF146
Ring finger protein 146
SEQ ID NOS: 11487-11498

RNF148
Ring finger protein 148
SEQ ID NOS: 11499-11500

RNF150
Ring finger protein 150
SEQ ID NOS: 11501-11505

RNF167
Ring finger protein 167
SEQ ID NOS: 11506-11516

RNF220
Ring finger protein 220
SEQ ID NOS: 11517-11523

RNF34
Ring finger protein 34, E3 ubiquitin protein ligase
SEQ ID NOS: 11524-11531

RNLS
Renalase, FAD-dependent amine oxidase
SEQ ID NOS: 11532-11534

RNPEP
Arginyl aminopeptidase (aminopeptidase B)
SEQ ID NOS: 11535-11540

ROR1
Receptor tyrosine kinase-like orphan receptor 1
SEQ ID NOS: 11541-11543

RPL3
Ribosomal protein L3
SEQ ID NOS: 11544-11549

RPLP2
Ribosomal protein, large, P2
SEQ ID NOS: 11550-11552

RPN2
Ribophorin II
SEQ ID NOS: 11553-11559

RPS27L
Ribosomal protein S27-like
SEQ ID NOS: 11560-11565

RS1
Retinoschisin 1
SEQ ID NO: 11566

RSF1
Remodeling and spacing factor 1
SEQ ID NOS: 11567-11573

RSPO1
R-spondin 1
SEQ ID NOS: 11574-11577

RSPO2
R-spondin 2
SEQ ID NOS: 11578-11585

RSPO3
R-spondin 3
SEQ ID NOS: 11586-11587

RSPO4
R-spondin 4
SEQ ID NOS: 11588-11589

RSPRY1
Ring finger and SPRY domain containing 1
SEQ ID NOS: 11590-11596

RTBDN
Retbindin
SEQ ID NOS: 11597-11609

RTN4RL1
Reticulon 4 receptor-like 1
SEQ ID NO: 11610

RTN4RL2
Reticulon 4 receptor-like 2
SEQ ID NOS: 11611-11613

SAA1
Serum amyloid A1
SEQ ID NOS: 11614-11616

SAA2
Serum amyloid A2
SEQ ID NOS: 11617-11622

SAA4
Serum amyloid A4, constitutive
SEQ ID NO: 11623

SAP30
Sin3A-associated protein, 30 kDa
SEQ ID NO: 11624

SAR1A
Secretion associated, Ras related GTPase 1A
SEQ ID NOS: 11625-11631

SARAF
Store-operated calcium entry-associated regulatory
SEQ ID NOS: 11632-11642

factor

SARM1
Sterile alpha and TIR motif containing 1
SEQ ID NOS: 11643-11646

SATB1
SATB homeobox 1
SEQ ID NOS: 11647-11659

SAXO2
Stabilizer of axonemal microtubules 2
SEQ ID NOS: 11660-11664

SBSN
Suprabasin
SEQ ID NOS: 11665-11667

SBSPON
Somatomedin B and thrombospondin, type 1
SEQ ID NO: 11668

domain containing

SCARF1
Scavenger receptor class F, member 1
SEQ ID NOS: 11669-11673

SCG2
Secretogranin II
SEQ ID NOS: 11674-11676

SCG3
Secretogranin III
SEQ ID NOS: 11677-11679

SCG5
Secretogranin V
SEQ ID NOS: 11680-11684

SCGB1A1
Secretoglobin, family 1A, member 1 (uteroglobin)
SEQ ID NOS: 11685-11686

SCGB1C1
Secretoglobin, family 1C, member 1
SEQ ID NO: 11687

SCGB1C2
Secretoglobin, family 1C, member 2
SEQ ID NO: 11688

SCGB1D1
Secretoglobin, family 1D, member 1
SEQ ID NO: 11689

SCGB1D2
Secretoglobin, family 1D, member 2
SEQ ID NO: 11690

SCGB1D4
Secretoglobin, family 1D, member 4
SEQ ID NO: 11691

SCGB2A1
Secretoglobin, family 2A, member 1
SEQ ID NO: 11692

SCGB2A2
Secretoglobin, family 2A, member 2
SEQ ID NOS: 11693-11694

SCGB2B2
Secretoglobin, family 2B, member 2
SEQ ID NOS: 11695-11696

SCGB3A1
Secretoglobin, family 3A, member 1
SEQ ID NO: 11697

SCGB3A2
Secretoglobin, family 3A, member 2
SEQ ID NOS: 11698-11699

SCN1B
Sodium channel, voltage gated, type I beta subunit
SEQ ID NOS: 11700-11705

SCN3B
Sodium channel, voltage gated, type III beta subunit
SEQ ID NOS: 11706-11710

SCPEP1
Serine carboxypeptidase 1
SEQ ID NOS: 11711-11718

SCRG1
Stimulator of chondrogenesis 1
SEQ ID NOS: 11719-11720

SCT
Secretin
SEQ ID NO: 11721

SCUBE1
Signal peptide, CUB domain, EGF-like 1
SEQ ID NOS: 11722-11725

SCUBE2
Signal peptide, CUB domain, EGF-like 2
SEQ ID NOS: 11726-11732

SCUBE3
Signal peptide, CUB domain, EGF-like 3
SEQ ID NO: 11733

SDC1
Syndecan 1
SEQ ID NOS: 11734-11738

SDF2
Stromal cell-derived factor 2
SEQ ID NOS: 11739-11741

SDF2L1
Stromal cell-derived factor 2-like 1
SEQ ID NO: 11742

SDF4
Stromal cell derived factor 4
SEQ ID NOS: 11743-11746

SDHAF2
Succinate dehydrogenase complex assembly factor 2
SEQ ID NOS: 11747-11754

SDHAF4
Succinate dehydrogenase complex assembly factor 4
SEQ ID NO: 11755

SDHB
Succinate dehydrogenase complex, subunit B, iron
SEQ ID NOS: 11756-11758

sulfur (Ip)

SDHD
Succinate dehydrogenase complex, subunit D,
SEQ ID NOS: 11759-11768

integral membrane protein

SEC14L3
SEC14-like lipid binding 3
SEQ ID NOS: 11769-11775

SEC16A
SEC16 homolog A, endoplasmic reticulum export
SEQ ID NOS: 11776-11782

factor

SEC16B
SEC16 homolog B, endoplasmic reticulum export
SEQ ID NOS: 11783-11786

factor

SEC22C
SEC22 homolog C, vesicle trafficking protein
SEQ ID NOS: 11787-11799

SEC31A
SEC31 homolog A, COPII coat complex component
SEQ ID NOS: 11800-11829

SECISBP2
SECIS binding protein 2
SEQ ID NOS: 11830-11834

SECTM1
Secreted and transmembrane 1
SEQ ID NOS: 11835-11842

SEL1L
Sel-1 suppressor of lin-12-like (C. elegans)
SEQ ID NOS: 11843-11845

SEPT15
15 kDa selenoprotein
SEQ ID NOS: 11846-11852

SELM
Selenoprotein M
SEQ ID NOS: 11853-11855

SEPN1
Selenoprotein N, 1
SEQ ID NOS: 11856-11859

SELO
Selenoprotein O
SEQ ID NOS: 11860-11861

SEPP1
Selenoprotein P, plasma, 1
SEQ ID NOS: 11862-11867

SEMA3A
Sema domain, immunoglobulin domain (Ig), short
SEQ ID NOS: 11868-11872

basic domain, secreted, (semaphorin) 3A

SEMA3B
Sema domain, immunoglobulin domain (Ig), short
SEQ ID NOS: 11873-11879

basic domain, secreted, (semaphorin) 3B

SEMA3C
Sema domain, immunoglobulin domain (Ig), short
SEQ ID NOS: 11880-11884

basic domain, secreted, (semaphorin) 3C

SEMA3E
Sema domain, immunoglobulin domain (Ig), short
SEQ ID NOS: 11885-11889

basic domain, secreted, (semaphorin) 3E

SEMA3F
Sema domain, immunoglobulin domain (Ig), short
SEQ ID NOS: 11890-11896

basic domain, secreted, (semaphorin) 3F

SEMA3G
Sema domain, immunoglobulin domain (Ig), short
SEQ ID NOS: 11897-11899

basic domain, secreted, (semaphorin) 3G

SEMA4A
Sema domain, immunoglobulin domain (Ig),
SEQ ID NOS: 11900-11908

transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4A

SEMA4B
Sema domain, immunoglobulin domain (Ig),
SEQ ID NOS: 11909-11919

transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4B

SEMA4C
Sema domain, immunoglobulin domain (Ig),
SEQ ID NOS: 11920-11922

transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4C

SEMA4D
Sema domain, immunoglobulin domain (Ig),
SEQ ID NOS: 11923-11936

transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4D

SEMA4F
Sema domain, immunoglobulin domain (Ig),
SEQ ID NOS: 11937-11945

transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4F

SEMA4G
Sema domain, immunoglobulin domain (Ig),
SEQ ID NOS: 11946-11953

transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4G

SEMA5A
Sema domain, seven thrombospondin repeats (type
SEQ ID NOS: 11954-11955

1 and type 1-like), transmembrane domain (TM) and

short cytoplasmic domain, (semaphorin) 5A

SEMA6A
Sema domain, transmembrane domain (TM), and
SEQ ID NOS: 11956-11963

cytoplasmic domain, (semaphorin) 6A

SEMA6C
Sema domain, transmembrane domain (TM), and
SEQ ID NOS: 11964-11969

cytoplasmic domain, (semaphorin) 6C

SEMA6D
Sema domain, transmembrane domain (TM), and
SEQ ID NOS: 11970-11983

cytoplasmic domain, (semaphorin) 6D

SEMG1
Semenogelin I
SEQ ID NO: 11984

SEMG2
Semenogelin II
SEQ ID NO: 11985

SEPT9
Septin 9
SEQ ID NOS: 11986-12022

SERPINA1
Serpin peptidase inhibitor, clade A (alpha-1
SEQ ID NOS: 12023-12039

antiproteinase, antitrypsin), member 1

SERPINA10
Serpin peptidase inhibitor, clade A (alpha-1
SEQ ID NOS: 12040-12043

antiproteinase, antitrypsin), member 10

SERPINA11
Serpin peptidase inhibitor, clade A (alpha-1
SEQ ID NO: 12044

antiproteinase, antitrypsin), member 11

SERPINA12
Serpin peptidase inhibitor, clade A (alpha-1
SEQ ID NOS: 12045-12046

antiproteinase, antitrypsin), member 12

SERPINA3
Serpin peptidase inhibitor, clade A (alpha-1
SEQ ID NOS: 12047-12053

antiproteinase, antitrypsin), member 3

SERPINA4
Serpin peptidase inhibitor, clade A (alpha-1
SEQ ID NOS: 12054-12056

antiproteinase, antitrypsin), member 4

SERPINA5
Serpin peptidase inhibitor, clade A (alpha-1
SEQ ID NOS: 12057-12068

antiproteinase, antitrypsin), member 5

SERPINA6
Serpin peptidase inhibitor, clade A (alpha-1
SEQ ID NOS: 12069-12071

antiproteinase, antitrypsin), member 6

SERPINA7
Serpin peptidase inhibitor, clade A (alpha-1
SEQ ID NOS: 12072-12073

antiproteinase, antitrypsin), member 7

SERPINA9
Serpin peptidase inhibitor, clade A (alpha-1
SEQ ID NOS: 12074-12080

antiproteinase, antitrypsin), member 9

SERPINB2
Serpin peptidase inhibitor, clade B (ovalbumin),
SEQ ID NOS: 12081-12085

member 2

SERPINC1
Serpin peptidase inhibitor, clade C (antithrombin),
SEQ ID NOS: 12086-12087

member 1

SERPIND1
Serpin peptidase inhibitor, clade D (heparin
SEQ ID NOS: 12088-12089

cofactor), member 1

SERPINE1
Serpin peptidase inhibitor, clade E (nexin,
SEQ ID NO: 12090

plasminogen activator inhibitor type 1), member 1

SERPINE2
Serpin peptidase inhibitor, clade E (nexin,
SEQ ID NOS: 12091-12097

plasminogen activator inhibitor type 1), member 2

SERPINE3
Serpin peptidase inhibitor, clade E (nexin,
SEQ ID NOS: 12098-12101

plasminogen activator inhibitor type 1), member 3

SERPINF1
Serpin peptidase inhibitor, clade F (alpha-2
SEQ ID NOS: 12102-12110

antiplasmin, pigment epithelium derived factor),

member 1

SERPINF2
Serpin peptidase inhibitor, clade F (alpha-2
SEQ ID NOS: 12111-12115

antiplasmin, pigment epithelium derived factor),

member 2

SERPING1
Serpin peptidase inhibitor, clade G (C1 inhibitor),
SEQ ID NOS: 12116-12126

member 1

SERPINH1
Serpin peptidase inhibitor, clade H (heat shock
SEQ ID NOS: 12127-12141

protein 47), member 1, (collagen binding protein 1)

SERPINI1
Serpin peptidase inhibitor, clade I (neuroserpin),
SEQ ID NOS: 12142-12146

member 1

SERPINI2
Serpin peptidase inhibitor, clade I (pancpin),
SEQ ID NOS: 12147-12153

member 2

SEZ6L2
Seizure related 6 homolog (mouse)-like 2
SEQ ID NOS: 12154-12160

SFRP1
Secreted frizzled-related protein 1
SEQ ID NOS: 12161-12162

SFRP2
Secreted frizzled-related protein 2
SEQ ID NO: 12163

SFRP4
Secreted frizzled-related protein 4
SEQ ID NOS: 12164-12165

SFRP5
Secreted frizzled-related protein 5
SEQ ID NO: 12166

SFTA2
Surfactant associated 2
SEQ ID NOS: 12167-12168

SFTPA1
Surfactant protein A1
SEQ ID NOS: 12169-12173

SFTPA2
Surfactant protein A2
SEQ ID NOS: 12174-12178

SFTPB
Surfactant protein B
SEQ ID NOS: 12179-12183

SFTPD
Surfactant protein D
SEQ ID NOS: 12184-12185

SFXN5
Sideroflexin 5
SEQ ID NOS: 12186-12190

SGCA
Sarcoglycan, alpha (50 kDa dystrophin-associated
SEQ ID NOS: 12191-12198

glycoprotein)

SGSH
N-sulfoglucosamine sulfohydrolase
SEQ ID NOS: 12199-12207

SH3RF3
SH3 domain containing ring finger 3
SEQ ID NO: 12208

SHBG
Sex hormone-binding globulin
SEQ ID NOS: 12209-12227

SHE
Src homology 2 domain containing E
SEQ ID NOS: 12228-12230

SHH
Sonic hedgehog
SEQ ID NOS: 12231-12234

SHKBP1
SH3KBP1 binding protein 1
SEQ ID NOS: 12235-12250

SIAE
Sialic acid acetylesterase
SEQ ID NOS: 12251-12253

SIDT2
SID1 transmembrane family, member 2
SEQ ID NOS: 12254-12263

SIGLEC10
Sialic acid binding Ig-like lectin 10
SEQ ID NOS: 12264-12272

SIGLEC6
Sialic acid binding Ig-like lectin 6
SEQ ID NOS: 12273-12278

SIGLEC7
Sialic acid binding Ig-like lectin 7
SEQ ID NOS: 12279-12283

SIGLECL1
SIGLEC family like 1
SEQ ID NOS: 12284-12289

SIGMAR1
Sigma non-opioid intracellular receptor 1
SEQ ID NOS: 12290-12293

SIL1
SIL1 nucleotide exchange factor
SEQ ID NOS: 12294-12302

SIRPB1
Signal-regulatory protein beta 1
SEQ ID NOS: 12303-12315

SIRPD
Signal-regulatory protein delta
SEQ ID NOS: 12316-12318

SLAMF1
Signaling lymphocytic activation molecule family
SEQ ID NOS: 12319-12321

member 1

SLAMF7
SLAM family member 7
SEQ ID NOS: 12322-12330

SLC10A3
Solute carrier family 10, member 3
SEQ ID NOS: 12331-12335

SLC15A3
Solute carrier family 15 (oligopeptide transporter),
SEQ ID NOS: 12336-12341

member 3

SLC25A14
Solute carrier family 25 (mitochondrial carrier,
SEQ ID NOS: 12342-12348

brain), member 14

SLC25A25
Solute carrier family 25 (mitochondrial carrier;
SEQ ID NOS: 12349-12355

phosphate carrier), member 25

SLC2A5
Solute carrier family 2 (facilitated glucose/fructose
SEQ ID NOS: 12356-12364

transporter), member 5

SLC35E3
Solute carrier family 35, member E3
SEQ ID NOS: 12365-12366

SLC39A10
Solute carrier family 39 (zinc transporter),
SEQ ID NOS: 12367-12373

member 10

SLC39A14
Solute carrier family 39 (zinc transporter),
SEQ ID NOS: 12374-12384

member 14

SLC39A4
Solute carrier family 39 (zinc transporter), member 4
SEQ ID NOS: 12385-12387

SLC39A5
Solute carrier family 39 (zinc transporter), member 5
SEQ ID NOS: 12388-12394

SLC3A1
Solute carrier family 3 (amino acid transporter heavy
SEQ ID NOS: 12395-12404

chain), member 1

SLC51A
Solute carrier family 51, alpha subunit
SEQ ID NOS: 12405-12409

SLC52A2
Solute carrier family 52 (riboflavin transporter),
SEQ ID NOS: 12410-12420

member 2

SLC5A6
Solute carrier family 5 (sodium/multivitamin and
SEQ ID NOS: 12421-12431

iodide cotransporter), member 6

SLC6A9
Solute carrier family 6 (neurotransmitter
SEQ ID NOS: 12432-12439

transporter, glycine), member 9

SLC8A1
Solute carrier family 8 (sodium/calcium exchanger),
SEQ ID NOS: 12440-12451

member 1

SLC8B1
Solute carrier family 8 (sodium/lithium/calcium
SEQ ID NOS: 12452-12462

exchanger), member B1

SLC9A6
Solute carrier family 9, subfamily A (NHE6, cation
SEQ ID NOS: 12463-12474

proton antiporter 6), member 6

SLCO1A2
Solute carrier organic anion transporter family,
SEQ ID NOS: 12475-12488

member 1A2

SLIT1
Slit guidance ligand 1
SEQ ID NOS: 12489-12492

SLIT2
Slit guidance ligand 2
SEQ ID NOS: 12493-12501

SLIT3
Slit guidance ligand 3
SEQ ID NOS: 12502-12504

SLITRK3
SLIT and NTRK-like family, member 3
SEQ ID NOS: 12505-12507

SLPI
Secretory leukocyte peptidase inhibitor
SEQ ID NO: 12508

SLTM
SAFB-like, transcription modulator
SEQ ID NOS: 12509-12522

SLURP1
Secreted LY6/PLAUR domain containing 1
SEQ ID NO: 12523

SMARCA2
SWI/SNF related, matrix associated, actin dependent
SEQ ID NOS: 12524-12571

regulator of chromatin, subfamily a, member 2

SMG6
SMG6 nonsense mediated mRNA decay factor
SEQ ID NOS: 12572-12583

SMIM7
Small integral membrane protein 7
SEQ ID NOS: 12584-12600

SMOC1
SPARC related modular calcium binding 1
SEQ ID NOS: 12601-12602

SMOC2
SPARC related modular calcium binding 2
SEQ ID NOS: 12603-12607

SMPDL3A
Sphingomyelin phosphodiesterase, acid-like 3A
SEQ ID NOS: 12608-12609

SMPDL3B
Sphingomyelin phosphodiesterase, acid-like 3B
SEQ ID NOS: 12610-12614

SMR3A
Submaxillary gland androgen regulated protein 3A
SEQ ID NO: 12615

SMR3B
Submaxillary gland androgen regulated protein 3B
SEQ ID NOS: 12616-12618

SNED1
Sushi, nidogen and EGF-like domains 1
SEQ ID NOS: 12619-12625

SNTB1
Syntrophin, beta 1 (dystrophin-associated protein
SEQ ID NOS: 12626-12628

A1, 59 kDa, basic component 1)

SNTB2
Syntrophin, beta 2 (dystrophin-associated protein
SEQ ID NOS: 12629-12633

A1, 59 kDa, basic component 2)

SNX14
Sorting nexin 14
SEQ ID NOS: 12634-12645

SOD3
Superoxide dismutase 3, extracellular
SEQ ID NOS: 12646-12647

SOST
Sclerostin
SEQ ID NO: 12648

SOSTDC1
Sclerostin domain containing 1
SEQ ID NOS: 12649-12650

SOWAHA
Sosondowah ankyrin repeat domain family member
SEQ ID NO: 12651

A

SPACA3
Sperm acrosome associated 3
SEQ ID NOS: 12652-12654

SPACA4
Sperm acrosome associated 4
SEQ ID NO: 12655

SPACA5
Sperm acrosome associated 5
SEQ ID NOS: 12656-12657

SPACA5B
Sperm acrosome associated 5B
SEQ ID NO: 12658

SPACA7
Sperm acrosome associated 7
SEQ ID NOS: 12659-12662

SPAG11A
Sperm associated antigen 11A
SEQ ID NOS: 12663-12671

SPAG11B
Sperm associated antigen 11B
SEQ ID NOS: 12672-12680

SPARC
Secreted protein, acidic, cysteine-rich (osteonectin)
SEQ ID NOS: 12681-12685

SPARCL1
SPARC-like 1 (hevin)
SEQ ID NOS: 12686-12695

SPATA20
Spermatogenesis associated 20
SEQ ID NOS: 12696-12709

SPESP1
Sperm equatorial segment protein 1
SEQ ID NO: 12710

SPINK1
Serine peptidase inhibitor, Kazal type 1
SEQ ID NOS: 12711-12712

SPINK13
Serine peptidase inhibitor, Kazal type 13 (putative)
SEQ ID NOS: 12713-12715

SPINK14
Serine peptidase inhibitor, Kazal type 14 (putative)
SEQ ID NOS: 12716-12717

SPINK2
Serine peptidase inhibitor, Kazal type 2 (acrosin-
SEQ ID NOS: 12718-12723

trypsin inhibitor)

SPINK4
Serine peptidase inhibitor, Kazal type 4
SEQ ID NOS: 12724-12725

SPINK5
Serine peptidase inhibitor, Kazal type 5
SEQ ID NOS: 12726-12731

SPINK6
Serine peptidase inhibitor, Kazal type 6
SEQ ID NOS: 12732-12734

SPINK7
Serine peptidase inhibitor, Kazal type 7 (putative)
SEQ ID NOS: 12735-12736

SPINK8
Serine peptidase inhibitor, Kazal type 8 (putative)
SEQ ID NO: 12737

SPINK9
Serine peptidase inhibitor, Kazal type 9
SEQ ID NOS: 12738-12739

SPINT1
Serine peptidase inhibitor, Kunitz type 1
SEQ ID NOS: 12740-12747

SPINT2
Serine peptidase inhibitor, Kunitz type, 2
SEQ ID NOS: 12748-12755

SPINT3
Serine peptidase inhibitor, Kunitz type, 3
SEQ ID NO: 12756

SPINT4
Serine peptidase inhibitor, Kunitz type 4
SEQ ID NO: 12757

SPOCK1
Sparc/osteonectin, cwcv and kazal-like domains
SEQ ID NOS: 12758-12761

proteoglycan (testican) 1

SPOCK2
Sparc/osteonectin, cwcv and kazal-like domains
SEQ ID NOS: 12762-12765

proteoglycan (testican) 2

SPOCK3
Sparc/osteonectin, cwcv and kazal-like domains
SEQ ID NOS: 12766-12791

proteoglycan (testican) 3

SPON1
Spondin 1, extracellular matrix protein
SEQ ID NO: 12792

SPON2
Spondin 2, extracellular matrix protein
SEQ ID NOS: 12793-12802

SPP1
Secreted phosphoprotein 1
SEQ ID NOS: 12803-12807

SPP2
Secreted phosphoprotein 2, 24 kDa
SEQ ID NOS: 12808-12810

SPRN
Shadow of prion protein homolog (zebrafish)
SEQ ID NO: 12811

SPRYD3
SPRY domain containing 3
SEQ ID NOS: 12812-12815

SPRYD4
SPRY domain containing 4
SEQ ID NO: 12816

SPTY2D1-
SPTY2D1 antisense RNA 1
SEQ ID NOS: 12817-12822

AS1

SPX
Spexin hormone
SEQ ID NOS: 12823-12824

SRGN
Serglycin
SEQ ID NO: 12825

SRL
Sarcalumenin
SEQ ID NOS: 12826-12828

SRP14
Signal recognition particle 14 kDa (homologous Alu
SEQ ID NOS: 12829-12832

RNA binding protein)

SRPX
Sushi-repeat containing protein, X-linked
SEQ ID NOS: 12833-12836

SRPX2
Sushi-repeat containing protein, X-linked 2
SEQ ID NOS: 12837-12840

SSC4D
Scavenger receptor cysteine rich family, 4 domains
SEQ ID NO: 12841

SSC5D
Scavenger receptor cysteine rich family, 5 domains
SEQ ID NOS: 12842-12845

SSPO
SCO-spondin
SEQ ID NO: 12846

SSR2
Signal sequence receptor, beta (translocon-
SEQ ID NOS: 12847-12856

associated protein beta)

SST
Somatostatin
SEQ ID NO: 12857

ST3GAL1
ST3 beta-galactoside alpha-2,3-sialyltransferase 1
SEQ ID NOS: 12858-12865

ST3GAL4
ST3 beta-galactoside alpha-2,3-sialyltransferase 4
SEQ ID NOS: 12866-12881

ST6GAL1
ST6 beta-galactosamide alpha-2,6-sialyltranferase 1
SEQ ID NOS: 12882-12897

ST6GALNAC
ST6 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-
SEQ ID NOS: 12898-12902

2
1,3)-N-acetylgalactosaminide alpha-2,6-

sialyltransferase 2

ST6GALNAC
ST6 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-
SEQ ID NOS: 12903-12904

5
1,3)-N-acetylgalactosaminide alpha-2,6-

sialyltransferase 5

ST6GALNAC
ST6 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-
SEQ ID NOS: 12905-12912

6
1,3)-N-acetylgalactosaminide alpha-2,6-

sialyltransferase 6

ST8SIA2
ST8 alpha-N-acetyl-neuraminide alpha-2,8-
SEQ ID NOS: 12913-12915

sialyltransferase 2

ST8SIA4
ST8 alpha-N-acetyl-neuraminide alpha-2,8-
SEQ ID NOS: 12916-12918

sialyltransferase 4

ST8SIA6
ST8 alpha-N-acetyl-neuraminide alpha-2,8-
SEQ ID NOS: 12919-12920

sialyltransferase 6

STARD7
StAR-related lipid transfer (START) domain
SEQ ID NOS: 12921-12922

containing 7

STATH
Statherin
SEQ ID NOS: 12923-12925

STC1
Stanniocalcin 1
SEQ ID NOS: 12926-12927

STC2
Stanniocalcin 2
SEQ ID NOS: 12928-12930

STMND1
Stathmin domain containing 1
SEQ ID NOS: 12931-12932

C7orf73
Chromosome 7 open reading frame 73
SEQ ID NOS: 12933-12934

STOML2
Stomatin (EPB72)-like 2
SEQ ID NOS: 12935-12938

STOX1
Storkhead box 1
SEQ ID NOS: 12939-12943

STRC
Stereocilin
SEQ ID NOS: 12944-12949

SUCLG1
Succinate-CoA ligase, alpha subunit
SEQ ID NOS: 12950-12951

SUDS3
SDS3 homolog, SIN3A corepressor complex
SEQ ID NO: 12952

component

SULF1
Sulfatase 1
SEQ ID NOS: 12953-12963

SULF2
Sulfatase 2
SEQ ID NOS: 12964-12968

SUMF1
Sulfatase modifying factor 1
SEQ ID NOS: 12969-12973

SUMF2
Sulfatase modifying factor 2
SEQ ID NOS: 12974-12987

SUSD1
Sushi domain containing 1
SEQ ID NOS: 12988-12993

SUSD5
Sushi domain containing 5
SEQ ID NOS: 12994-12995

SVEP1
Sushi, von Willebrand factor type A, EGF and
SEQ ID NOS: 12996-12998

pentraxin domain containing 1

SWSAP1
SWIM-type zinc finger 7 associated protein 1
SEQ ID NO: 12999

SYAP1
Synapse associated protein 1
SEQ ID NO: 13000

SYCN
Syncollin
SEQ ID NO: 13001

TAC1
Tachykinin, precursor 1
SEQ ID NOS: 13002-13004

TAC3
Tachykinin 3
SEQ ID NOS: 13005-13014

TAC4
Tachykinin 4 (hemokinin)
SEQ ID NOS: 13015-13020

TAGLN2
Transgelin 2
SEQ ID NOS: 13021-13024

TAPBP
TAP binding protein (tapasin)
SEQ ID NOS: 13025-13030

TAPBPL
TAP binding protein-like
SEQ ID NOS: 13031-13032

TBL2
Transducin (beta)-like 2
SEQ ID NOS: 13033-13045

TBX10
T-box 10
SEQ ID NO: 13046

TCF12
Transcription factor 12
SEQ ID NOS: 13047-13060

TCN1
Transcobalamin I (vitamin B12 binding protein, R
SEQ ID NO: 13061

binder family)

TCN2
Transcobalamin II
SEQ ID NOS: 13062-13065

TCTN1
Tectonic family member 1
SEQ ID NOS: 13066-13084

TCTN3
Tectonic family member 3
SEQ ID NOS: 13085-13089

TDP2
Tyrosyl-DNA phosphodiesterase 2
SEQ ID NOS: 13090-13091

C14orf80
Chromosome 14 open reading frame 80
SEQ ID NOS: 13092-13105

TEK
TEK tyrosine kinase, endothelial
SEQ ID NOS: 13106-13110

TEPP
Testis, prostate and placenta expressed
SEQ ID NOS: 13111-13112

TEX101
Testis expressed 101
SEQ ID NOS: 13113-13114

TEX264
Testis expressed 264
SEQ ID NOS: 13115-13126

C1orf234
Chromosome 1 open reading frame 234
SEQ ID NOS: 13127-13129

TF
Transferrin
SEQ ID NOS: 13130-13136

TFAM
Transcription factor A, mitochondrial
SEQ ID NOS: 13137-13139

TFF1
Trefoil factor 1
SEQ ID NO: 13140

TFF2
Trefoil factor 2
SEQ ID NO: 13141

TFF3
Trefoil factor 3 (intestinal)
SEQ ID NOS: 13142-13144

TFPI
Tissue factor pathway inhibitor (lipoprotein-
SEQ ID NOS: 13145-13154

associated coagulation inhibitor)

TFPI2
Tissue factor pathway inhibitor 2
SEQ ID NOS: 13155-13156

TG
Thyroglobulin
SEQ ID NOS: 13157-13166

TGFB1
Transforming growth factor, beta 1
SEQ ID NOS: 13167-13168

TGFB2
Transforming growth factor, beta 2
SEQ ID NOS: 13169-13170

TGFB3
Transforming growth factor, beta 3
SEQ ID NOS: 13171-13172

TGFBI
Transforming growth factor, beta-induced, 68 kDa
SEQ ID NOS: 13173-13180

TGFBR1
Transforming growth factor, beta receptor 1
SEQ ID NOS: 13181-13190

TGFBR3
Transforming growth factor, beta receptor III
SEQ ID NOS: 13191-13197

THBS1
Thrombospondin 1
SEQ ID NOS: 13198-13199

THBS2
Thrombospondin 2
SEQ ID NOS: 13200-13202

THBS3
Thrombospondin 3
SEQ ID NOS: 13203-13207

THBS4
Thrombospondin 4
SEQ ID NOS: 13208-13209

THOC3
THO complex 3
SEQ ID NOS: 13210-13219

THPO
Thrombopoietin
SEQ ID NOS: 13220-13225

THSD4
Thrombospondin, type I, domain containing 4
SEQ ID NOS: 13226-13229

THY1
Thy-1 cell surface antigen
SEQ ID NOS: 13230-13235

TIE1
Tyrosine kinase with immunoglobulin-like and EGF-
SEQ ID NOS: 13236-13237

like domains 1

TIMMDC1
Translocase of inner mitochondrial membrane
SEQ ID NOS: 13238-13245

domain containing 1

TIMP1
TIMP metallopeptidase inhibitor 1
SEQ ID NOS: 13246-13250

TIMP2
TIMP metallopeptidase inhibitor 2
SEQ ID NOS: 13251-13255

TIMP3
TIMP metallopeptidase inhibitor 3
SEQ ID NO: 13256

TIMP4
TIMP metallopeptidase inhibitor 4
SEQ ID NO: 13257

TINAGL1
Tubulointerstitial nephritis antigen-like 1
SEQ ID NOS: 13258-13260

TINF2
TERF1 (TRF1)-interacting nuclear factor 2
SEQ ID NOS: 13261-13270

TLL2
Tolloid-like 2
SEQ ID NO: 13271

TLR1
Toll-like receptor 1
SEQ ID NOS: 13272-13277

TLR3
Toll-like receptor 3
SEQ ID NOS: 13278-13280

TM2D2
TM2 domain containing 2
SEQ ID NOS: 13281-13286

TM2D3
TM2 domain containing 3
SEQ ID NOS: 13287-13294

TM7SF3
Transmembrane 7 superfamily member 3
SEQ ID NOS: 13295-13309

TM95F1
Transmembrane 9 superfamily member 1
SEQ ID NOS: 13310-13320

TMCO6
Transmembrane and coiled-coil domains 6
SEQ ID NOS: 13321-13328

TMED1
Transmembrane p24 trafficking protein 1
SEQ ID NOS: 13329-13335

TMED2
Transmembrane p24 trafficking protein 2
SEQ ID NOS: 13336-13338

TMED3
Transmembrane p24 trafficking protein 3
SEQ ID NOS: 13339-13342

TMED4
Transmembrane p24 trafficking protein 4
SEQ ID NOS: 13343-13345

TMED5
Transmembrane p24 trafficking protein 5
SEQ ID NOS: 13346-13349

TMED7
Transmembrane p24 trafficking protein 7
SEQ ID NOS: 13350-13351

TMED7-
TMED7-TICAM2 readthrough
SEQ ID NOS: 13352-13353

TICAM2

TMEM108
Transmembrane protein 108
SEQ ID NOS: 13354-13362

TMEM116
Transmembrane protein 116
SEQ ID NOS: 13363-13374

TMEM119
Transmembrane protein 119
SEQ ID NOS: 13375-13378

TMEM155
Transmembrane protein 155
SEQ ID NOS: 13379-13382

TMEM168
Transmembrane protein 168
SEQ ID NOS: 13383-13388

TMEM178A
Transmembrane protein 178A
SEQ ID NOS: 13389-13390

TMEM179
Transmembrane protein 179
SEQ ID NOS: 13391-13396

TMEM196
Transmembrane protein 196
SEQ ID NOS: 13397-13401

TMEM199
Transmembrane protein 199
SEQ ID NOS: 13402-13405

TMEM205
Transmembrane protein 205
SEQ ID NOS: 13406-13419

TMEM213
Transmembrane protein 213
SEQ ID NOS: 13420-13423

TMEM25
Transmembrane protein 25
SEQ ID NOS: 13424-13440

TMEM30C
Transmembrane protein 30C
SEQ ID NO: 13441

TMEM38B
Transmembrane protein 38B
SEQ ID NOS: 13442-13446

TMEM44
Transmembrane protein 44
SEQ ID NOS: 13447-13456

TMEM52
Transmembrane protein 52
SEQ ID NOS: 13457-13461

TMEM52B
Transmembrane protein 52B
SEQ ID NOS: 13462-13464

TMEM59
Transmembrane protein 59
SEQ ID NOS: 13465-13472

TMEM67
Transmembrane protein 67
SEQ ID NOS: 13473-13484

TMEM70
Transmembrane protein 70
SEQ ID NOS: 13485-13487

TMEM87A
Transmembrane protein 87A
SEQ ID NOS: 13488-13497

TMEM94
Transmembrane protein 94
SEQ ID NOS: 13498-13513

TMEM95
Transmembrane protein 95
SEQ ID NOS: 13514-13516

TMIGD1
Transmembrane and immunoglobulin domain
SEQ ID NOS: 13517-13518

containing 1

TMPRSS12
Transmembrane (C-terminal) protease, serine 12
SEQ ID NOS: 13519-13520

TMPRSS5
Transmembrane protease, serine 5
SEQ ID NOS: 13521-13532

TMUB1
Transmembrane and ubiquitin-like domain
SEQ ID NOS: 13533-13539

containing 1

TMX2
Thioredoxin-related transmembrane protein 2
SEQ ID NOS: 13540-13547

TMX3
Thioredoxin-related transmembrane protein 3
SEQ ID NOS: 13548-13555

TNC
Tenascin C
SEQ ID NOS: 13556-13564

TNFAIP6
Tumor necrosis factor, alpha-induced protein 6
SEQ ID NO: 13565

TNFRSF11A
Tumor necrosis factor receptor superfamily,
SEQ ID NOS: 13566-13570

member 11a, NFKB activator

TNFRSF11B
Tumor necrosis factor receptor superfamily,
SEQ ID NOS: 13571-13572

member 11b

TNFRSF12A
Tumor necrosis factor receptor superfamily,
SEQ ID NOS: 13573-13578

member 12A

TNFRSF14
Tumor necrosis factor receptor superfamily,
SEQ ID NOS: 13579-13585

member 14

TNFRSF18
Tumor necrosis factor receptor superfamily,
SEQ ID NOS: 13586-13589

member 18

TNFRSF1A
Tumor necrosis factor receptor superfamily,
SEQ ID NOS: 13590-13598

member 1A

TNFRSF1B
Tumor necrosis factor receptor superfamily,
SEQ ID NOS: 13599-13600

member 1B

TNFRSF25
Tumor necrosis factor receptor superfamily,
SEQ ID NOS: 13601-13612

member 25

TNFRSF6B
Tumor necrosis factor receptor superfamily,
SEQ ID NO: 13613

member 6b, decoy

TNFSF11
Tumor necrosis factor (ligand) superfamily,
SEQ ID NOS: 13614-13618

member 11

TNFSF12
Tumor necrosis factor (ligand) superfamily,
SEQ ID NOS: 13619-13620

member 12

TNFSF12-
TNFSF12-TNFSF13 readthrough
SEQ ID NO: 13621

TNFSF13

TNFSF15
Tumor necrosis factor (ligand) superfamily,
SEQ ID NOS: 13622-13623

member 15

TNN
Tenascin N
SEQ ID NOS: 13624-13626

TNR
Tenascin R
SEQ ID NOS: 13627-13629

TNXB
Tenascin XB
SEQ ID NOS: 13630-13636

FAM179B
Family with sequence similarity 179, member B
SEQ ID NOS: 13637-13642

TOMM7
Translocase of outer mitochondrial membrane 7
SEQ ID NOS: 13643-13646

homolog (yeast)

TOP1MT
Topoisomerase (DNA) I, mitochondrial
SEQ ID NOS: 13647-13661

TOR1A
Torsin family 1, member A (torsin A)
SEQ ID NO: 13662

TOR1B
Torsin family 1, member B (torsin B)
SEQ ID NOS: 13663-13664

TOR2A
Torsin family 2, member A
SEQ ID NOS: 13665-13671

TOR3A
Torsin family 3, member A
SEQ ID NOS: 13672-13676

TPD52
Tumor protein D52
SEQ ID NOS: 13677-13689

TPO
Thyroid peroxidase
SEQ ID NOS: 13690-13700

TPP1
Tripeptidyl peptidase I
SEQ ID NOS: 13701-13718

TPSAB1
Tryptase alpha/beta 1
SEQ ID NOS: 13719-13721

TPSB2
Tryptase beta 2 (gene/pseudogene)
SEQ ID NOS: 13722-13724

TPSD1
Tryptase delta 1
SEQ ID NOS: 13725-13726

TPST1
Tyrosylprotein sulfotransferase 1
SEQ ID NOS: 13727-13729

TPST2
Tyrosylprotein sulfotransferase 2
SEQ ID NOS: 13730-13738

TRABD2A
TraB domain containing 2A
SEQ ID NOS: 13739-13741

TRABD2B
TraB domain containing 2B
SEQ ID NO: 13742

TREH
Trehalase (brush-border membrane glycoprotein)
SEQ ID NOS: 13743-13745

TREM1
Triggering receptor expressed on myeloid cells 1
SEQ ID NOS: 13746-13749

TREM2
Triggering receptor expressed on myeloid cells 2
SEQ ID NOS: 13750-13752

TRH
Thyrotropin-releasing hormone
SEQ ID NOS: 13753-13754

TRIM24
Tripartite motif containing 24
SEQ ID NOS: 13755-13756

TRIM28
Tripartite motif containing 28
SEQ ID NOS: 13757-13762

TRIO
Trio Rho guanine nucleotide exchange factor
SEQ ID NOS: 13763-13769

TRNP1
TMF1-regulated nuclear protein 1
SEQ ID NOS: 13770-13771

TSC22D4
TSC22 domain family, member 4
SEQ ID NOS: 13772-13775

TSHB
Thyroid stimulating hormone, beta
SEQ ID NOS: 13776-13777

TSHR
Thyroid stimulating hormone receptor
SEQ ID NOS: 13778-13785

TSKU
Tsukushi, small leucine rich proteoglycan
SEQ ID NOS: 13786-13790

TSLP
Thymic stromal lymphopoietin
SEQ ID NOS: 13791-13793

TSPAN3
Tetraspanin 3
SEQ ID NOS: 13794-13799

TSPAN31
Tetraspanin 31
SEQ ID NOS: 13800-13806

TSPEAR
Thrombospondin-type laminin G domain and EAR
SEQ ID NOS: 13807-13810

repeats

TTC13
Tetratricopeptide repeat domain 13
SEQ ID NOS: 13811-13817

TTC19
Tetratricopeptide repeat domain 19
SEQ ID NOS: 13818-13823

TTC9B
Tetratricopeptide repeat domain 9B
SEQ ID NO: 13824

TTLL11
Tubulin tyrosine ligase-like family member 11
SEQ ID NOS: 13825-13829

TTR
Transthyretin
SEQ ID NOS: 13830-13832

TWSG1
Twisted gastrulation BMP signaling modulator 1
SEQ ID NOS: 13833-13835

TXNDC12
Thioredoxin domain containing 12 (endoplasmic
SEQ ID NOS: 13836-13838

reticulum)

TXNDC15
Thioredoxin domain containing 15
SEQ ID NOS: 13839-13845

TXNDC5
Thioredoxin domain containing 5 (endoplasmic
SEQ ID NOS: 13846-13847

reticulum)

TXNRD2
Thioredoxin reductase 2
SEQ ID NOS: 13848-13860

TYRP1
Tyrosinase-related protein 1
SEQ ID NOS: 13861-13863

UBAC2
UBA domain containing 2
SEQ ID NOS: 13864-13868

UBALD1
UBA-like domain containing 1
SEQ ID NOS: 13869-13877

UBAP2
Ubiquitin associated protein 2
SEQ ID NOS: 13878-13884

UBXN8
UBX domain protein 8
SEQ ID NOS: 13885-13891

UCMA
Upper zone of growth plate and cartilage matrix
SEQ ID NOS: 13892-13893

associated

UCN
Urocortin
SEQ ID NO: 13894

UCN2
Urocortin 2
SEQ ID NO: 13895

UCN3
Urocortin 3
SEQ ID NO: 13896

UGGT2
UDP-glucose glycoprotein glucosyltransferase 2
SEQ ID NOS: 13897-13902

UGT1A10
UDP glucuronosyltransferase 1 family, polypeptide
SEQ ID NOS: 13903-13904

A10

UGT2A1
UDP glucuronosyltransferase 2 family, polypeptide
SEQ ID NOS: 13905-13909

A1, complex locus

UGT2B11
UDP glucuronosyltransferase 2 family, polypeptide
SEQ ID NO: 13910

B11

UGT2B28
UDP glucuronosyltransferase 2 family, polypeptide
SEQ ID NOS: 13911-13912

B28

UGT2B4
UDP glucuronosyltransferase 2 family, polypeptide
SEQ ID NOS: 13913-13916

B4

UGT2B7
UDP glucuronosyltransferase 2 family, polypeptide
SEQ ID NOS: 13917-13920

B7

UGT3A1
UDP glycosyltransferase 3 family, polypeptide A1
SEQ ID NOS: 13921-13926

UGT3A2
UDP glycosyltransferase 3 family, polypeptide A2
SEQ ID NOS: 13927-13930

UGT8
UDP glycosyltransferase 8
SEQ ID NOS: 13931-13933

ULBP3
UL16 binding protein 3
SEQ ID NOS: 13934-13935

UMOD
Uromodulin
SEQ ID NOS: 13936-13947

UNC5C
Unc-5 netrin receptor C
SEQ ID NOS: 13948-13952

UPK3B
Uroplakin 3B
SEQ ID NOS: 13953-13955

USP11
Ubiquitin specific peptidase 11
SEQ ID NOS: 13956-13959

USP14
Ubiquitin specific peptidase 14 (tRNA-guanine
SEQ ID NOS: 13960-13966

transglycosylase)

USP3
Ubiquitin specific peptidase 3
SEQ ID NOS: 13967-13982

CIRH1A
Cirrhosis, autosomal recessive 1A (cirhin)
SEQ ID NOS: 13983-13992

UTS2
Urotensin 2
SEQ ID NOS: 13993-13995

UTS2B
Urotensin 2B
SEQ ID NOS: 13996-14001

UTY
Ubiquitously transcribed tetratricopeptide repeat
SEQ ID NOS: 14002-14014

containing, Y-linked

UXS1
UDP-glucuronate decarboxylase 1
SEQ ID NOS: 14015-14022

VASH1
Vasohibin 1
SEQ ID NOS: 14023-14025

VCAN
Versican
SEQ ID NOS: 14026-14032

VEGFA
Vascular endothelial growth factor A
SEQ ID NOS: 14033-14058

VEGFB
Vascular endothelial growth factor B
SEQ ID NOS: 14059-14061

VEGFC
Vascular endothelial growth factor C
SEQ ID NO: 14062

FIGF
C-fos induced growth factor (vascular endothelial
SEQ ID NO: 14063

growth factor D)

VGF
VGF nerve growth factor inducible
SEQ ID NOS: 14064-14066

VIP
Vasoactive intestinal peptide
SEQ ID NOS: 14067-14069

VIPR2
Vasoactive intestinal peptide receptor 2
SEQ ID NOS: 14070-14073

VIT
Vitrin
SEQ ID NOS: 14074-14081

VKORC1
Vitamin K epoxide reductase complex, subunit 1
SEQ ID NOS: 14082-14089

VLDLR
Very low density lipoprotein receptor
SEQ ID NOS: 14090-14092

VMO1
Vitelline membrane outer layer 1 homolog (chicken)
SEQ ID NOS: 14093-14096

VNN1
Vanin 1
SEQ ID NO: 14097

VNN2
Vanin 2
SEQ ID NOS: 14098-14111

VNN3
Vanin 3
SEQ ID NOS: 14112-14123

VOPP1
Vesicular, overexpressed in cancer, prosurvival
SEQ ID NOS: 14124-14136

protein 1

VPREB1
Pre-B lymphocyte 1
SEQ ID NOS: 14137-14138

VPREB3
Pre-B lymphocyte 3
SEQ ID NOS: 14139-14140

VPS37B
Vacuolar protein sorting 37 homolog B (S. cerevisiae)
SEQ ID NOS: 14141-14143

VPS51
Vacuolar protein sorting 51 homolog (S. cerevisiae)
SEQ ID NOS: 14144-14155

VSIG1
V-set and immunoglobulin domain containing 1
SEQ ID NOS: 14156-14158

VSIG10
V-set and immunoglobulin domain containing 10
SEQ ID NOS: 14159-14160

VSTM1
V-set and transmembrane domain containing 1
SEQ ID NOS: 14161-14167

VSTM2A
V-set and transmembrane domain containing 2A
SEQ ID NOS: 14168-14171

VSTM2B
V-set and transmembrane domain containing 2B
SEQ ID NO: 14172

VSTM2L
V-set and transmembrane domain containing 2 like
SEQ ID NOS: 14173-14175

VSTM4
V-set and transmembrane domain containing 4
SEQ ID NOS: 14176-14177

VTN
Vitronectin
SEQ ID NOS: 14178-14179

VWA1
Von Willebrand factor A domain containing 1
SEQ ID NOS: 14180-14183

VWA2
Von Willebrand factor A domain containing 2
SEQ ID NOS: 14184-14185

VWA5B2
Von Willebrand factor A domain containing 5B2
SEQ ID NOS: 14186-14187

VWA7
Von Willebrand factor A domain containing 7
SEQ ID NO: 14188

VWC2
Von Willebrand factor C domain containing 2
SEQ ID NO: 14189

VWC2L
Von Willebrand factor C domain containing protein
SEQ ID NOS: 14190-14191

2-like

VWCE
Von Willebrand factor C and EGF domains
SEQ ID NOS: 14192-14196

VWDE
Von Willebrand factor D and EGF domains
SEQ ID NOS: 14197-14202

VWF
Von Willebrand factor
SEQ ID NOS: 14203-14205

WDR25
WD repeat domain 25
SEQ ID NOS: 14206-14212

WDR81
WD repeat domain 81
SEQ ID NOS: 14213-14222

WDR90
WD repeat domain 90
SEQ ID NOS: 14223-14230

WFDC1
WAP four-disulfide core domain 1
SEQ ID NOS: 14231-14233

WFDC10A
WAP four-disulfide core domain 10A
SEQ ID NO: 14234

WFDC10B
WAP four-disulfide core domain 10B
SEQ ID NOS: 14235-14236

WFDC11
WAP four-disulfide core domain 11
SEQ ID NOS: 14237-14239

WFDC12
WAP four-disulfide core domain 12
SEQ ID NO: 14240

WFDC13
WAP four-disulfide core domain 13
SEQ ID NO: 14241

WFDC2
WAP four-disulfide core domain 2
SEQ ID NOS: 14242-14246

WFDC3
WAP four-disulfide core domain 3
SEQ ID NOS: 14247-14250

WFDC5
WAP four-disulfide core domain 5
SEQ ID NOS: 14251-14252

WFDC6
WAP four-disulfide core domain 6
SEQ ID NOS: 14253-14254

WFDC8
WAP four-disulfide core domain 8
SEQ ID NOS: 14255-14256

WFIKKN1
WAP, follistatin/kazal, immunoglobulin, kunitz and
SEQ ID NO: 14257

netrin domain containing 1

WFIKKN2
WAP, follistatin/kazal, immunoglobulin, kunitz and
SEQ ID NOS: 14258-14259

netrin domain containing 2

DFNB31
Deafness, autosomal recessive 31
SEQ ID NOS: 14260-14263

WIF1
WNT inhibitory factor 1
SEQ ID NOS: 14264-14266

WISP1
WNT1 inducible signaling pathway protein 1
SEQ ID NOS: 14267-14271

WISP2
WNT1 inducible signaling pathway protein 2
SEQ ID NOS: 14272-14274

WISP3
WNT1 inducible signaling pathway protein 3
SEQ ID NOS: 14275-14282

WNK1
WNK lysine deficient protein kinase 1
SEQ ID NOS: 14283-14296

WNT1
Wingless-type MMTV integration site family,
SEQ ID NOS: 14297-14298

member 1

WNT10B
Wingless-type MMTV integration site family,
SEQ ID NOS: 14299-14303

member 10B

WNT11
Wingless-type MMTV integration site family,
SEQ ID NOS: 14304-14306

member 11

WNT16
Wingless-type MMTV integration site family,
SEQ ID NOS: 14307-14308

member 16

WNT2
Wingless-type MMTV integration site family
SEQ ID NOS: 14309-14311

member 2

WNT3
Wingless-type MMTV integration site family,
SEQ ID NO: 14312

member 3

WNT3A
Wingless-type MMTV integration site family,
SEQ ID NO: 14313

member 3A

WNT5A
Wingless-type MMTV integration site family,
SEQ ID NOS: 14314-14317

member 5A

WNT5B
Wingless-type MMTV integration site family,
SEQ ID NOS: 14318-14324

member 5B

WNT6
Wingless-type MMTV integration site family,
SEQ ID NO: 14325

member 6

WNT7A
Wingless-type MMTV integration site family,
SEQ ID NO: 14326

member 7A

WNT7B
Wingless-type MMTV integration site family,
SEQ ID NOS: 14327-14331

member 7B

WNT8A
Wingless-type MMTV integration site family,
SEQ ID NOS: 14332-14335

member 8A

WNT8B
Wingless-type MMTV integration site family,
SEQ ID NO: 14336

member 8B

WNT9A
Wingless-type MMTV integration site family,
SEQ ID NO: 14337

member 9A

WNT9B
Wingless-type MMTV integration site family,
SEQ ID NOS: 14338-14340

member 9B

WSB1
WD repeat and SOCS box containing 1
SEQ ID NOS: 14341-14350

WSCD1
WSC domain containing 1
SEQ ID NOS: 14351-14360

WSCD2
WSC domain containing 2
SEQ ID NOS: 14361-14364

XCL1
Chemokine (C motif) ligand 1
SEQ ID NO: 14365

XCL2
Chemokine (C motif) ligand 2
SEQ ID NO: 14366

XPNPEP2
X-prolyl aminopeptidase (aminopeptidase P) 2,
SEQ ID NOS: 14367-14368

membrane-bound

XXYLT1
Xyloside xylosyltransferase 1
SEQ ID NOS: 14369-14374

XYLT1
Xylosyltransferase I
SEQ ID NO: 14375

XYLT2
Xylosyltransferase II
SEQ ID NOS: 14376-14381

ZFYVE21
Zinc finger, FYVE domain containing 21
SEQ ID NOS: 14382-14386

ZG16
Zymogen granule protein 16
SEQ ID NO: 14387

ZG16B
Zymogen granule protein 16B
SEQ ID NOS: 14388-14391

ZIC4
Zic family member 4
SEQ ID NOS: 14392-14400

ZNF207
Zinc finger protein 207
SEQ ID NOS: 14401-14411

ZNF26
Zinc finger protein 26
SEQ ID NOS: 14412-14415

ZNF34
Zinc finger protein 34
SEQ ID NOS: 14416-14419

ZNF419
Zinc finger protein 419
SEQ ID NOS: 14420-14434

ZNF433
Zinc finger protein 433
SEQ ID NOS: 14435-14444

ZNF449
Zinc finger protein 449
SEQ ID NOS: 14445-14446

ZNF488
Zinc finger protein 488
SEQ ID NOS: 14447-14448

ZNF511
Zinc finger protein 511
SEQ ID NOS: 14449-14450

ZNF570
Zinc finger protein 570
SEQ ID NOS: 14451-14456

ZNF691
Zinc finger protein 691
SEQ ID NOS: 14457-14464

ZNF98
Zinc finger protein 98
SEQ ID NOS: 14465-14468

ZPBP
Zona pellucida binding protein
SEQ ID NOS: 14469-14472

ZPBP2
Zona pellucida binding protein 2
SEQ ID NOS: 14473-14476

ZSCAN29
Zinc finger and SCAN domain containing 29
SEQ ID NOS: 14477-14483

In certain embodiments, the therapeutic protein is not secreted, but rather functions intracellularly.

In certain embodiments, the therapeutic protein is not secreted, but rather directs a modified cell of the disclosure to a cell niche of a subject's body.

In certain embodiments of the methods of the disclosure, the subject has a disease or disorder and the plurality of therapeutic immune cells or immune precursor cells improves a sign or symptom of the disease or disorder, optionally by providing a therapeutic protein systemically or locally within the subject that acts upon the immune cell, the immune precursor cell or a second cell in the subject. Exemplary therapeutic secreted proteins may be used as a monotherapy or in combination with another therapy in the treatment or prevention of any disease or disorder. These secreted proteins may be used as a monotherapy or in combination with another therapy for enzyme replacement and/or administration of biologic therapeutics.

Inducible Proapoptotic Polypeptides

Inducible proapoptotic polypeptides of the disclosure are superior to existing inducible polypeptides because the inducible proapoptotic polypeptides of the disclosure are far less immunogenic. While inducible proapoptotic polypeptides of the disclosure are recombinant polypeptides, and, therefore, non-naturally occurring, the sequences that are recombined to produce the inducible proapoptotic polypeptides of the disclosure do not comprise non-human sequences that the host human immune system could recognize as “non-self” and, consequently, induce an immune response in the subject receiving an inducible proapoptotic polypeptide of the disclosure, a cell comprising the inducible proapoptotic polypeptide or a composition comprising the inducible proapoptotic polypeptide or the cell comprising the inducible proapoptotic polypeptide.

Modified cells and/or transposons of the disclosure may comprise an inducible proapoptotic polypeptide comprising (a) a ligand binding region, (b) a linker, and (c) a proapoptotic polypeptide, wherein the inducible proapoptotic polypeptide does not comprise a non-human sequence. In certain embodiments, the non-human sequence comprises a restriction site. In certain embodiments, the ligand binding region may be a multimeric ligand binding region. Inducible proapoptotic polypeptides of the disclosure may also be referred to as an “iC9 safety switch”. In certain embodiments, modified cells and/or transposons of the disclosure may comprise an inducible caspase polypeptide comprising (a) a ligand binding region, (b) a linker, and (c) a caspase polypeptide, wherein the inducible proapoptotic polypeptide does not comprise a non-human sequence. In certain embodiments, modified cells and/or transposons of the disclosure may comprise an inducible caspase polypeptide comprising (a) a ligand binding region, (b) a linker, and (c) a caspase polypeptide, wherein the inducible proapoptotic polypeptide does not comprise a non-human sequence. In certain embodiments, transposons of the disclosure may comprise an inducible caspase polypeptide comprising (a) a ligand binding region, (b) a linker, and (c) a truncated caspase 9 polypeptide, wherein the inducible proapoptotic polypeptide does not comprise a non-human sequence. In certain embodiments of the inducible proapoptotic polypeptides, inducible caspase polypeptides or truncated caspase 9 polypeptides of the disclosure, the ligand binding region may comprise a FK506 binding protein 12 (FKBP12) polypeptide. In certain embodiments, the amino acid sequence of the ligand binding region that comprise a FK506 binding protein 12 (FKBP12) polypeptide may comprise a modification at position 36 of the sequence. The modification may be a substitution of valine (V) for phenylalanine (F) at position 36 (F36V).

In certain embodiments, the FKBP12 polypeptide is encoded by an amino acid sequence comprising

(SEQ ID NO: 14635)

GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKPFKF

MLGKQEVIRGWEEGVAQMSVGQRAKLTISPDVAYGATGHPGIIPPHAT

LVFDVELLKLE.

In certain embodiments, the FKBP12 polypeptide is encoded by a nucleic acid sequence comprising

(SEQ ID NO: 14636)

GGGGTCCAGGTCGAGACTATTTCACCAGGGGATGGGCGAACATTTCCA

AAAAGGGGCCAGACTTGCGTCGTGCATTACACCGGGATGCTGGAGGAC

GGGAAGAAAGTGGACAGCTCCAGGGATCGCAACAAGCCCTTCAAGTTC

ATGCTGGGAAAGCAGGAAGTGATCCGAGGATGGGAGGAAGGCGTGGCA

CAGATGTCAGTCGGCCAGCGGGCCAAACTGACCATTAGCCCTGACTAC

GCTTATGGAGCAACAGGCCACCCAGGGATCATTCCCCCTCATGCCACC

CTGGTCTTCGATGTGGAACTGCTGAAGCTGGAG.

In certain embodiments, the induction agent specific for the ligand binding region may comprise a FK506 binding protein 12 (FKBP12) polypeptide having a substitution of valine (V) for phenylalanine (F) at position 36 (F36V) comprises AP20187 and/or AP1903, both synthetic drugs.

In certain embodiments of the inducible proapoptotic polypeptides, inducible caspase polypeptides or truncated caspase 9 polypeptides of the disclosure, the linker region is encoded by an amino acid comprising GGGGS (SEQ ID NO: 14637) or a nucleic acid sequence comprising GGAGGAGGAGGATCC (SEQ ID NO: 14638). In certain embodiments, the nucleic acid sequence encoding the linker does not comprise a restriction site.

In certain embodiments of the truncated caspase 9 polypeptides of the disclosure, the truncated caspase 9 polypeptide is encoded by an amino acid sequence that does not comprise an arginine (R) at position 87 of the sequence. Alternatively, or in addition, in certain embodiments of the inducible proapoptotic polypeptides, inducible caspase polypeptides or truncated caspase 9 polypeptides of the disclosure, the truncated caspase 9 polypeptide is encoded by an amino acid sequence that does not comprise an alanine (A) at position 282 the sequence. In certain embodiments of the inducible proapoptotic polypeptides, inducible caspase polypeptides or truncated caspase 9 polypeptides of the disclosure, the truncated caspase 9 polypeptide is encoded by an amino acid comprising

(SEQ ID NO: 14639)

GFGDVGALESLRGNADLAYILSMEPCGHCLIINNVNFCRESGLRTRTG

SNIDCEKLRRRFSSLHFMVEVKGDLTAKKMVLALLELAQQDHGALDCC

VVVILSHGCQASHLQFPGAVYGTDGCPVSVEKIVNIFNGTSCPSLGGK

PKLFFIQACGGEQKDHGFEVASTSPEDESPGSNPEPDATPFQEGLRTF

DQLDAISSLPTPSDIFVSYSTFPGFVSWRDPKSGSWYVETLDDIFEQW

AHSEDLQSLLLRVANAVSVKGIYKQMPGCFNFLRKKLFFKTS

or a nucleic acid sequence comprising

(SEQ ID NO: 14640)

TTTGGGGACGTGGGGGCCCTGGAGTCTCTGCGAGGAAATGCCGATCTG

GCTTACATCCTGAGCATGGAACCCTGCGGCCACTGTCTGATCATTAAC

AATGTGAACTTCTGCAGAGAAAGCGGACTGCGAACACGGACTGGCTCC

AATATTGACTGTGAGAAGCTGCGGAGAAGGTTCTCTAGTCTGCACTTT

ATGGTCGAAGTGAAAGGGGATCTGACCGCCAAGAAAATGGTGCTGGCC

CTGCTGGAGCTGGCTCAGCAGGACCATGGAGCTCTGGATTGCTGCGTG

GTCGTGATCCTGTCCCACGGGTGCCAGGCTTCTCATCTGCAGTTCCCC

GGAGCAGTGTACGGAACAGACGGCTGTCCTGTCAGCGTGGAGAAGATC

GTCAACATCTTCAACGGCACTTCTTGCCCTAGTCTGGGGGGAAAGCCA

AAACTGTTCTTTATCCAGGCCTGTGGCGGGGAACAGAAAGATCACGGC

TTCGAGGTGGCCAGCACCAGCCCTGAGGACGAATCACCAGGGAGCAAC

CCTGAACCAGATGCAACTCCATTCCAGGAGGGACTGAGGACCTTTGAC

CAGCTGGATGCTATCTCAAGCCTGCCCACTCCTAGTGACATTTTCGTG

TCTTACAGTACCTTCCCAGGCTTTGTCTCATGGCGCGATCCCAAGTCA

GGGAGCTGGTACGTGGAGACACTGGACGACATCTTTGAACAGTGGGCC

CATTCAGAGGACCTGCAGAGCCTGCTGCTGCGAGTGGCAAACGCTGTC

TCTGTGAAGGGCATCTACAAACAGATGCCCGGGTGCTTCAATTTTCTG

AGAAAGAAACTGTTCTTTAAGACTTCC.

In certain embodiments of the inducible proapoptotic polypeptides, wherein the polypeptide comprises a truncated caspase 9 polypeptide, the inducible proapoptotic polypeptide is encoded by an amino acid sequence comprising

(SEQ ID NO: 14641)

GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKPFKF

MLGKQEVIRGWEEGVAQMSVGQRAKLTISPDVAYGATGHPGIIPPHAT

LVFDVELLKLEGGGGSGFGDVGALESLRGNADLAYILSMEPCGHCLII

NNVNFCRESGLRTRTGSNIDCEKLRRRFSSLHFMVEVKGDLTAKKMVL

ALLELAQQDHGALDCCVVVILSHGCQASHLQFPGAVYGTDGCPVSVEK

IVNIFNGTSCPSLGGKPKLFFIQACGGEQKDHGFEVASTSPEDESPGS

NPEPDATPFQEGLRTFDQLDAIS SLPTP SDIFVSYSTFPGFVSWRD

PKSGSWYVETLDDIFEQWAHSEDLQSLLLRVANAVSVKGIYKQMPGCF

NFLRKKLFFKTS

or the nucleic acid sequence comprising

(SEQ ID NO: 14642)

ggggtccaggtcgagactatttcaccaggggatgggcgaacatttcca

aaaaggggccagacttgcgtcgtgcattacaccgggatgctggaggac

gggaagaaagtggacagctccagggatcgcaacaagcccttcaagttc

atgctgggaaagcaggaagtgatccgaggatgggaggaaggcgtggca

cagatgtcagtcggccagcgggccaaactgaccattagccctgactac

gcttatggagcaacaggccacccagggatcattccccctcatgccacc

ctggtcttcgatgtggaactgctgaagctggagggaggaggaggatcc

ggatttggggacgtgggggccctggagtctctgcgaggaaatgccgat

ctggcttacatcctgagcatggaaccctgcggccactgtctgatcatt

aacaatgtgaacttctgcagagaaagcggactgcgaacacggactggc

tccaatattgactgtgagaagctgcggagaaggttctctagtctgcac

tttatggtcgaagtgaaaggggatctgaccgccaagaaaatggtgctg

gccctgctggagctggctcagcaggaccatggagctctggattgctgc

gtggtcgtgatcctgtcccacgggtgccaggcttctcatctgcagttc

cccggagcagtgtacggaacagacggctgtcctgtcagcgtggagaag

atcgtcaacatcttcaacggcacttcttgccctagtctggggggaaag

ccaaaactgttctttatccaggcctgtggcggggaacagaaagatcac

ggcttcgaggtggccagcaccagccctgaggacgaatcaccagggagc

aaccctgaaccagatgcaactccattccaggagggactgaggaccttt

gaccagctggatgctatctcaagcctgcccactcctagtgacattttc

gtgtcttacagtaccttcccaggctttgtctcatggcgcgatcccaag

tcagggagctggtacgtggagacactggacgacatctttgaacagtgg

gcccattcagaggacctgcagagcctgctgctgcgagtggcaaacgct

gtctctgtgaagggcatctacaaacagatgcccgggtgcttcaattac

tgagaaagaaactgttctttaagacttcc.

Construct Elements

Transposons and other delivery vectors of the disclosure may comprise at least one self-cleaving peptide(s) located, for example, between one or more of a sequence encoding an inducible proapoptotic polypeptide of the disclosure, a sequence encoding a therapeutic protein of the disclosure and a selection gene of the disclosure.

Transposons and other delivery vectorsof the disclosure may comprise at least two self-cleaving peptide(s), a first self-cleaving peptide located, for example, upstream or immediately upstream of an inducible proapoptotic polypeptide of the disclosure of the disclosure and a second first self-cleaving peptide located, for example, downstream or immediately upstream of an inducible proapoptotic polypeptide of the disclosure of the disclosure.

The at least one self-cleaving peptide may comprise, for example, a T2A peptide, GSG-T2A peptide, an E2A peptide, a GSG-E2A peptide, an F2A peptide, a GSG-F2A peptide, a P2A peptide, or a GSG-P2A peptide. A T2A peptide may comprise an amino acid sequence comprising EGRGSLLTCGDVEENPGP (SEQ ID NO: 14643) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising EGRGSLLTCGDVEENPGP (SEQ ID NO: 14643). A GSG-T2A peptide may comprise an amino acid sequence comprising GSGEGRGSLLTCGDVEENPGP (SEQ ID NO: 14644) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGEGRGSLLTCGDVEENPGP (SEQ ID NO: 14644). A GSG-T2A peptide may comprise a nucleic acid sequence comprising

(SEQ ID NO: 14645)

ggatctggagagggaaggggaagcctgctgacctgtggagacgtggagg

aaaacccaggacca.

An E2A peptide may comprise an amino acid sequence comprising QCTNYALLKLAGDVESNPGP (SEQ ID NO: 14646) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising QCTNYALLKLAGDVESNPGP (SEQ ID NO: 14646). A GSG-E2A peptide may comprise an amino acid sequence comprising GSGQCTNYALLKLAGDVESNPGP (SEQ ID NO: 14647) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGQCTNYALLKLAGDVESNPGP (SEQ ID NO: 14647). An F2A peptide may comprise an amino acid sequence comprising VKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14648) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising VKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14648). A GSG-F2A peptide may comprise an amino acid sequence comprising GSGVKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14649) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGVKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14649). A P2A peptide may comprise an amino acid sequence comprising ATNFSLLKQAGDVEENPGP (SEQ ID NO: 14650) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising ATNFSLLKQAGDVEENPGP (SEQ ID NO: 14650). A GSG-P2A peptide may comprise an amino acid sequence comprising GSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 14651) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 14651).

Transposons and other delivery vectors of the disclosure may comprise a first and a second self-cleaving peptide, the first self-cleaving peptide located, for example, upstream of one or more of a sequence encoding a therapeutic protein of the disclosure the second self-cleaving peptide located, for example, downstream of a sequence encoding a therapeutic protein of the disclosure. The first and/or the second self-cleaving peptide may comprise, for example, a T2A peptide, GSG-T2A peptide, an E2A peptide, a GSG-E2A peptide, an F2A peptide, a GSG-F2A peptide, a P2A peptide, or a GSG-P2A peptide. A T2A peptide may comprise an amino acid sequence comprising EGRGSLLTCGDVEENPGP (SEQ ID NO: 14643) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising EGRGSLLTCGDVEENPGP (SEQ ID NO: 14643). A GSG-T2A peptide may comprise an amino acid sequence comprising GSGEGRGSLLTCGDVEENPGP (SEQ ID NO: 14644) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGEGRGSLLTCGDVEENPGP (SEQ ID NO: 14644). A GSG-T2A peptide may comprise a nucleic acid sequence comprising

(SEQ ID NO: 14645)

ggatctggagagggaaggggaagcctgctgacctgtggagacgtggagg

aaaacccaggacca.

Transposons of the disclosure may comprise a selection gene. The selection gene may encode a gene product essential for cell viability and survival. The selection gene may encode a gene product essential for cell viability and survival when challenged by selective cell culture conditions. Selective cell culture conditions may comprise a compound harmful to cell viability or survival and wherein the gene product confers resistance to the compound.

By “stable transformation” is intended that the polynucleotide construct introduced into a cell integrates into the genome of the host and is capable of being inherited by progeny thereof.

By “transient transformation” is intended that a polynucleotide construct introduced into the host does not integrate into the genome of the host.

All percentages and ratios are calculated based on the total composition unless otherwise indicated.

Every maximum numerical limitation given throughout this disclosure includes every lower numerical limitation, as if such lower numerical limitations were expressly written herein. Every minimum numerical limitation given throughout this disclosure will include every higher numerical limitation, as if such higher numerical limitations were expressly written herein. Every numerical range given throughout this disclosure will include every narrower numerical range that falls within such broader numerical range, as if such narrower numerical ranges were all expressly written herein.

The values disclosed herein are not to be understood as being strictly limited to the exact numerical values recited. Instead, unless otherwise specified, each such value is intended to mean both the recited value and a functionally equivalent range surrounding that value. For example, a value disclosed as “20 μm” is intended to mean “about 20 μm.”

Every document cited herein, including any cross referenced or related patent or application, is hereby incorporated herein by reference in its entirety unless expressly excluded or otherwise limited. The citation of any document is not an admission that it is prior art with respect to any invention disclosed or claimed herein or that it alone, or in any combination with any other reference or references, teaches, suggests or discloses any such invention. Further, to the extent that any meaning or definition of a term in this document conflicts with any meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to that term in this document shall govern.

While particular embodiments of the disclosure have been illustrated and described, various other changes and modifications can be made without departing from the spirit and scope of the disclosure. The scope of the appended claims includes all such changes and modifications that are within the scope of this disclosure.

EXAMPLES

In order that the invention disclosed herein may be more efficiently understood, examples are provided below. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting the invention in any manner. Throughout these examples, molecular cloning reactions, and other standard recombinant DNA techniques, were carried out according to methods described in Maniatis et al., Molecular Cloning—A Laboratory Manual, 2nd ed., Cold Spring Harbor Press (1989), using commercially available reagents, except where otherwise noted.

Example 1: Ex Vivo Genetic Modification of T Cells

The piggyBac™ (PB) transposon system was used for genetically modifying human lymphocytes for production of autologous CAR-T immunotherapies and other applications. T Lymphocytes purified from patient blood or apheresis product was electroporated with a plasmid DNA transposon and a transposase. Several different electroporation systems have been used for T cell delivery of the transposon system, including the Neon (Thermo Fisher), BTX ECM 830 (Harvard Apparatus), Gene Pulser (BioRad), MaxCyte PulseAgile (MaxCyte), and the Amaxa 2B and Amaxa 4D (Lonza). Some were tested using manufacturer provided or recommended electroporation buffer, as well as several in-house developed buffers. Results were consistent with the prevailing dogma that resting T lymphocytes are particularly refractory to DNA transfection and that there appeared to be an inverse relationship between electroporation efficiency, as measured by GFP expression from the electroporated plasmid, and cell viability. FIG. 1 shows an example of an experiment testing multiple electroporation systems and nucleofection programs.

To further test whether or not plasmid DNA was toxic to T cells during nucleofection, primary human T lymphocytes were electroporated with two different DNA plasmids. The first plasmid was a pmaxGFP™ plasmid that is provided as a control plasmid in the Lonza Amaxa nucleofection kit. It is highly purified by HPLC and does not contain endotoxin at detectable levels. The second plasmid was our in-house produced PB transposon encoding a human EF1 alpha promoter driving GFP. Transfection efficiency, as measured by GFP expression from the electroporated plasmid, and cell viability was assessed by FACS at days 2, 3, and 6 post-electroporation. Data are displayed in FIG. 2. While mock electroporated cells (no plasmid DNA) exhibited relatively high levels of cell viability by day 6 post-electroporation, 54%, T cells electroporated with either plasmid were only 1.4-2.6% viable. These data show that plasmid DNA was cytotoxic to T lymphocytes. In addition, these data show that DNA-mediated toxicity was not due to transposon element such as the ITR regions or the core insulators since the pmaxGFP™ plasmid are devoid of these elements and was also cytotoxic at the same DNA concentration. Both plasmids are approximately the same size, meaning that similar amounts of DNA were electroporated into the T cells.

To test whether or not DNA-mediated toxicity in T cells was dose dependent, we performed a titration of our PB-GFP plasmid. FIG. 3 shows that as the dose of plasmid DNA added to the nucleofection reaction was increased incrementally (1.3, 2.5, 5.0, 10.0, and 20.0 μg of plasmid DNA), cell viability decreased as measured at both day 1 and 5 post-nucleofection. Even 1.3 μg of plasmid DNA was responsible for a 2.4-fold decrease in T cell viability by day 4.

Since it was clear that plasmid DNA is toxic to T cells during nucleofection, we considered whether or not extracellular plasmid DNA was contributing to cell death. FIG. 4 shows that extracellular plasmid DNA was not cytotoxic to T cells. In that experiment, 5 μg of plasmid DNA was added to the cells 45 min post-electroporation and little cell death was observed at day 1 or day 4. Similarly, when 5 μg of plasmid DNA was added to the nucleofection reaction in the absence of electroporation, little cell death was observed. However, when the plasmid DNA was added before the electroporation reaction, the cells exhibited a 2.0-fold reduction in cell viability at day 1 and a 13.2-fold reduction at day 4.

Since DNA-mediated toxicity is dose dependent, we next focused our attention on ways to reduce the total amount of DNA delivered to the T cells that is required for transposition. One relatively straightforward way of achieving this would be to deliver the transposase as encoded in mRNA instead of encoded in DNA. mRNA delivery to primary human T cells is very efficient, resulting in high transfection efficiency and high viability. We subcloned the Super piggyBac™ (SPB) transposase enzyme into our in-house mRNA production vector and produced high quality SPB mRNA. Co-delivery of PB-GFP transposon with various doses of SPB mRNA (30, 10, 3.3, 3, 1, 0.33 μg mRNA) in Jurkat cells demonstrated strong transposition at all doses tested (FIG. 5). These data show that SPB transposase can be delivered and are equally effective as either plasmid DNA or mRNA. In addition, that the amount of SPB mRNA makes little difference in overall transposition efficiency in Jurkats, in either overall percentage of GFP+ cells or in the MFI of GFP expression. To see if this also holds true for T lymphocytes, we delivered PB-GFP with either SPB plasmid DNA, at a 3:1 ratio, or 5 μg of SPB mRNA. Seven (7) days following the nucleofection reaction and the addition of IL7 and IL15, GFP transposition was assessed. FIG. 6 shows that SPB mRNA efficiently mediated transposition of the GFP transposon into T lymphocytes. Importantly, T cell viability was improved when co-delivering the SPB as an mRNA as opposed to a pDNA; 32.4% versus 25.4%, respectively. These data suggest that co-delivery of SPB as mRNA would be dose-sparing in the total amount of plasmid DNA being delivered to T cells and is thus less cytotoxic.

Since the current plasmid transposon also contains a backbone required for plasmid amplification in bacteria, it is possible to significantly reduce the total amount of DNA by excluding this sequence. This may be achieved by restriction digest of the plasmid transposon prior to the nucleofection reaction. In addition, this could be achieved by administering the transposon as a PCR product or as a Doggybone™ DNA, which is a double stranded DNA that is produced in vitro by a mechanism that excludes the initial backbone elements required for bacterial replication of the plasmid.

We performed a pilot experiment to see whether or not plasmid transposon needed to be circular, or if it could be delivered to the cell in a linear fashion. To test this, transposon was incubated overnight with a restriction enzyme (ApaLI) to linearize the plasmid. Either uncut or linearized plasmid is electroporated into primary T lymphocytes. GFP expression was assessed 2 days later. FIG. 7 shows that linearized plasmid was also efficiently delivered to the cell nucleus. These data demonstrate that linear transposon products can also be efficiently electroporated into primary human T cells.

We show above that plasmid DNA is toxic in primary T lymphocytes, but we have observed that this toxic effect is not as dramatic in tumor cell lines and other transformed cells. Based upon this observation, we hypothesized that primary T lymphocytes may be refractory to plasmid DNA transfection due to heightened DNA sensing pathways, which would protect immune cells from infection by viruses and bacteria. If these data are a result of heightened DNA sensing mechanisms, then it may be possible to enhance plasmid transfection efficiency and/or cell viability by the addition of DNA sensing pathway inhibitors to the post-nucleofection reaction. Thus, we tested a number of different reagents that inhibited the TLR-9 pathway, caspase pathway, or those involved in cytoplasmic double stranded DNA sensing. These reagents include Bafilomycin Al, which is an autophagy inhibitor that interferes with endosomal acidification and blocks NFkB signaling by TLR9, Chloroquine, which is a TLR9 antagonist, Quinacrine, which is a TLR9 antagonist and a cGAS antagonist, AC-YVAD-CMK, which is a caspase 1 inhibitor targeting the AIM2 pathway, Z-VAD-FMK, which is a pan caspase inhibitor, Z-IETD-FMK, which is a caspase 8 inhibitor triggered by the TLR9 pathway. In addition, we also tested the stimulation of electroporated T cells by the addition of the cytokines IL7 and IL15, as well as the addition of anti-CD3 anti-CD28 Dynabeads® Human T-Expander CD3/CD28 beads. Results are displayed in FIG. 8. We found that few of the compounds or caspase inhibitors had any positive effect on cell viability at day 4 post-nucleofection at the doses tested. However, we acknowledge that further dosing studies may be required to better test these reagents. It may also be more effective to inhibit these pathways genetically. Two post-nucleofection conditions did enhance viability of the T cells. The addition of IL7 and IL15, whether they were added either 1 hour or 1 day following electroporation, enhanced viability over 3-fold when compared with introduction of the plasmid transposon alone without additional treatment. Furthermore, stimulation of the T cells post-nucleofection using either activator or expander beads also dramatically enhanced T cell viability; stimulation was better when the beads were added 1 hour or 1 day post-nucleofection as compared to adding the beads 2 days post. Lastly, we also tested ROCK inhibitor and the removal of dead cells from the culture using the Dead Cell Removal kit from Miltenyi, but saw no improvement in cell viability.

To further expand upon these findings demonstrating that stimulation of the T cells post-nucleofection improves viability, we repeated the study using the addition of the cytokine IL7 and IL15. FIG. 9 shows that the addition of these cytokines each at a dose of 20 ng/mL either immediately following nucleofection or up to 1 hour post enhanced cell viability up to 2.9-fold when compared to no treatment. Addition of these cytokines up to 1 day post-nucleofection also enhanced viability, but not as strong as the prior time points.

Since we found that immediate stimulation of the T cells post-nucleofection was able to increase cell viability, we hypothesized that stimulating the cells prior to nucleofection may also enhance viability and transfection efficiency. To test this, we stimulated primary T lymphocytes either 2, 3, or 4 days prior to transposon nucleofection. FIG. 10 shows that some level of transposition occurs when the transposon and the transposase are co-delivered after the T cells have been stimulated prior to the nucleofection reaction. The efficacy of pre-stimulation may be influenced by the kinetics of stimulation and may therefore be dependent upon the precise type of expander technology chosen.

Example 2: Ex Vivo Genetic Modification of NK Cells

The piggyBac™ (PB) transposon system was used for genetically modifying human NK cells. Non-activated NK cells derived from CD3-depleted leukopheresis (containing CD14/CD19/CD56+ cells) were were electroporated with plasmid piggyBac transposon DNA encoding GFP and mRNA encoding Super piggyBac transposase using the program indicated in FIG. 14 from Lonza 4D nucleofector or BTX ECM 830 (500V, 700 usec pulse length, 0.2 mm electrode gap, one pulse). Transposed cells were co-cultured (stimulated) at day 2 with artificial antigen presenting cells (aAPCs). Fluorescent activated cell sorting (FACS) analysis of GFP percent at day 7 post-EP (day 5 post-stimulation) is shown in FIG. 14. Percent viability is the percentage of 7-Aminoactinomycin (7AAD)-negative cells at day 2 post-EP.

Transposition of non-activated NK cells from CD3-depleted leukopheresis (containing CD14/CD19/CD56+ cells) is shown in FIG. 15. Cells were electroporated with a plasmid piggyBac transposon encoding GFP and 5 ug mRNA encoding Super piggyBac transposase using the indicated Maxcyte electroporator program. Transposed cells were stimulated at day 2 with artificial antigen presenting cells (aAPCs). FACS plots (FIG. 15A) and a bar graph (FIG. 15B) from the analysis of percent GFP+ of CD56+ cells at day 6 post-EP and day 4 post-stimulation are shown. Percent viability is the percentage of 7AAD-negative cells at day 2 post EP.

FIG. 16 shows that there is dose-dependent DNA-mediated cytotoxicity in NK cells. FACS analysis of live cells (7AAD-ve/FSC, or Forward Scatter) at day 2 post-EP using Lonza 4D Nucleofector program DN-100. FACS plots (FIG. 16A) are quantified in graph (FIG. 16B). 5x10E6 cells were electroporated per electroporation in 100 uL P3 buffer in cuvettes. Cells were electroporated with no DNA (Mock) or varying amounts of piggyBac GFP transposon co-delivered with 5 ug super piggyBac mRNA.

Example 3: In Vitro Differentiation of piggyBac Modified HSPCs into B Cells

Human CD34+ HSPCs were electroporated with mRNA encoding Super piggyBac along with a piggyBac transposon encoding GFP. After electroporation, HSPCs were primed for B cell differentiation in presence of human IL-3, Flt3L, TPO, SCF, and G-CSF for 5 days. On day 6, cells were transferred to a layer of MS-5 feeder cells and fed bi-weekly, along with transfer to a fresh layer of feeders once per week. On day 34 of the in vitro differentiation process, CD19+B cells were generated and detectable in the culture (FIG. 17). A fraction of the B cells were positive for the GFP piggyBac transgene (FIG. 17, lower right panel) demonstrating that the piggyBac DNA Modification System can be used to modify HSPCs, which can then be later differentiated into more differentiated immune cell types. This technique allows for the derivation of genetically-modified immune cells from hematopoietic progenitors.

Number	Date	Country
62608546	Dec 2017	US
62558286	Sep 2017	US
62552861	Aug 2017	US

TRANSPOSON SYSTEM AND METHODS OF USE

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