COMPOSITIONS AND METHODS FOR INHIBITING KETOHEXOKINASE (KHK)

Abstract

Oligonucleotides are provided herein that inhibit KHK expression. Also provided are compositions including the same and uses thereof, particularly uses relating to treating diseases, disorders and/or conditions associated with KHK expression.

Description

SEQUENCE LISTING

The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Mar. 30, 2022, is named 02-0559-US-3_SL.txt and is 398,673 bytes in size.

FIELD OF THE INVENTION

The invention relates to oligonucleotides that inhibit KHK expression, compositions including the same and uses thereof. The invention also relates to methods for treating diseases, disorders and/or conditions associated with KHK expression.

BACKGROUND OF THE INVENTION

Ketohexokinase (KHK) is an important enzyme in fructose metabolism. KHK catalyzes the conversion of D-fructose to fructose-1-phosphate. Under conditions of elevated fructose consumption, a major part of fructose-1 phosphate contributes to fatty-acid and triglyceride synthesis among other things. In the liver, uncontrolled regulation of this process can lead to diseases such as non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). Similarly, fructose metabolism converts fructose to glucose in the liver. Increased levels of glucose can lead to glucose intolerance (i.e., pre-diabetes, type-2 diabetes, and impaired fasting glucose). Excess glucose is converted to fatty acids and triglycerides and heightens the risk of developing cardiovascular disease (e.g., hypertension). Decreasing the amount of KHK in the liver is likely to reduce the development of, or symptoms of these diseases. Strategies for targeting the KHK gene to prevent such diseases are needed. RNAi agents targeting the KHK gene have been disclosed e.g., in WO 2015/123264 and WO 2020/060986.

SUMMARY OF THE INVENTION

The disclosure is based in part on the discovery that oligonucleotides (e.g., RNAi oligonucleotides) reduce KHK expression in the liver. Specifically, target sequences within KHK mRNA were identified and oligonucleotides that bind to these target sequences and inhibit KHK mRNA expression were generated. As demonstrated herein, the oligonucleotides inhibited murine KHK expression, and/or monkey and human KHK expression in the liver. Without being bound by theory, the oligonucleotides described herein are useful for treating a disease, disorder or condition associated with KHK expression (e.g., Non-alcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH)). In some embodiments, the oligonucleotides described herein are useful for treating a disease, disorder or condition associated with mutations in the KHK gene.

Accordingly, in some aspects, the present disclosure provides a double stranded RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising an antisense strand and a sense strand, wherein the antisense strand and the sense strand form a duplex region, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387 and wherein the region of complementarity is at least 15 contiguous nucleotides in length, or a pharmaceutically acceptable salt thereof. In some aspects, the present disclosure provides a double stranded RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising an antisense strand and a sense strand, wherein the antisense strand and the sense strand form a duplex region, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, wherein the region of complementarity is at least 15 contiguous nucleotides in length, and wherein KHK expression is reduced by at least 50%.

In any of the foregoing or related aspects, the sense strand comprises a sequence set forth in any one of SEQ ID NOs: 4-387.

In any of the foregoing or related aspects, the anti-sense strand comprises a sequence set forth in any one of SEQ ID NOs: 388-771.

In other aspects, the disclosure provides a double stranded RNAi oligonucleotide for inhibiting expression of KHK, wherein said double stranded RNAi agent comprises a sense strand and an antisense strand forming a duplex region, wherein said sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the nucleotide sequences of SEQ ID NO: 4-387 and said antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequences of SEQ ID NO: 388-771, or a pharmaceutically acceptable salt thereof. In other aspects, the disclosure provides a double stranded RNAi oligonucleotide for inhibiting expression of KHK, wherein said double stranded RNAi agent comprises a sense strand and an antisense strand forming a duplex region, wherein said sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the nucleotide sequences of SEQ ID NO:4-387 and said antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequences of SEQ ID NO: 388-771, and wherein KHK expression is reduced by at least 50%, or a pharmaceutically acceptable salt thereof.

In any of the foregoing or related aspects, the sense strand is 15 to 50 nucleotides in length. In some aspects, the sense strand is 18 to 36 nucleotides in length. In other aspects, the sense strand is 15 to 30 nucleotides in length. In some aspects, the antisense strand is 15-30 nucleotides in length. In some aspects, the antisense strand is 22 nucleotides in length.

In any of the foregoing or related aspects, the antisense strand and the sense strand form a duplex region of at least 19 nucleotides in length. In any of the foregoing or related aspects, the antisense strand and the sense strand form a duple region of at least 20 nucleotides in length. In any of the foregoing or related aspects, the antisense strand and the sense strand form a duplex region of 20 nucleotides in length. In some aspects, the antisense strand is 22 nucleotides in length and the antisense strand and the sense strand form a duplex region of at least 19 nucleotides in length. In some aspects, the antisense strand is 22 nucleotides in length and the antisense strand and the sense strand form a duplex region of at least 20 nucleotides in length. In some aspects, the antisense strand is 22 nucleotides in length and the antisense strand and the sense strand form a duplex region of 20 nucleotides in length.

In any of the foregoing or related aspects, the antisense strand comprises a region of complementarity of at least 19 contiguous nucleotides in length, optionally at least 20 nucleotides in length.

In any of the foregoing or related aspects, the sense strand comprises at its 3′ end a stem-loop set forth as: S1-L-S2, wherein S1 is complementary to S2, and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length.

In some aspects, the disclosure provides an RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising a sense strand of 15 to 50 nucleotides in length and an antisense strand, wherein the sense strand and the antisense strand form a duplex region, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is at least 15 contiguous nucleotides in length, or a pharmaceutically acceptable salt thereof.

In yet other aspects, the disclosure provides an RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising a sense strand of 15 to 50 nucleotides in length and an antisense strand of 15 to 30 nucleotides in length, wherein the sense strand and the antisense strand form a duplex region, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is at least 15 contiguous nucleotides in length, or a pharmaceutically acceptable salt thereof.

In other aspects, the disclosure provides an RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising a sense strand of 15 to 50 nucleotides in length and an antisense strand, wherein the sense strand and the antisense strand form a duplex region, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is 19 contiguous nucleotides in length, optionally 20 nucleotides in length, or a pharmaceutically acceptable salt thereof.

In other aspects, the disclosure provides an RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising a sense strand of 18 to 36 nucleotides in length and an antisense strand, wherein the sense strand and the antisense strand form a duplex region, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is 19 contiguous nucleotides in length, optionally 20 nucleotides in length, or a pharmaceutically acceptable salt thereof.

In yet other aspects, the disclosure provides an RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising a sense strand of 18 to 36 nucleotides in length and an antisense strand of 22 nucleotides in length, wherein the sense strand and the antisense strand form a duplex region, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is 19 contiguous nucleotides in length, optionally 20 nucleotides in length, or a pharmaceutically acceptable salt thereof.

In some aspects, the disclosure provides an RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising a sense strand of 18 to 36 nucleotides in length and an antisense strand of 22 nucleotides in length, wherein the sense strand and the antisense strand form a duplex region, wherein the 3′ end of the sense strand comprises a stem-loop set forth as S1-L-S2, wherein S1 is complementary to S2, and wherein L forms a loop between S1 and S2 of 3-5 nucleotides in length, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is 19 contiguous nucleotides in length, optionally 20 nucleotides in length, or a pharmaceutically acceptable salt thereof.

In other aspects, the disclosure provides an RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising a sense strand of 36 nucleotides in length and an antisense strand of 22 nucleotides in length, wherein the sense strand and the antisense strand form a duplex region, wherein the 3′ end of the sense strand comprises a stem-loop set forth as S1-L-S2, wherein S1 is complementary to S2, and wherein L forms a loop between S1 and S2 of 3-5 nucleotides in length, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is 19 contiguous nucleotides in length, optionally 20 nucleotides in length, or a pharmaceutically acceptable salt thereof.

In yet other aspects, the disclosure provides an RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising a sense strand of 36 nucleotides in length and an antisense strand of 22 nucleotides in length, wherein the sense strand and the antisense strand form a duplex region of at least 19 nucleotides in length, optionally 20 nucleotides in length, wherein the 3′ end of the sense strand comprises a stem-loop set forth as S1-L-S2, wherein S1 is complementary to S2, and wherein L forms a loop between S1 and S2 of 3-5 nucleotides in length, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is 19 contiguous nucleotides in length, optionally 20 nucleotides in length, or a pharmaceutically acceptable salt thereof.

In some aspects, the disclosure provides a RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising:

• • (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is selected from SEQ ID NOs: 948-953; and
  • (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand,

wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand. In some aspects, the RNAi oligonucleotide comprises a stem-loop at the 3′ terminus, wherein the stem loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide comprising a stem loop set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length. In some aspects, L is a triloop or a tetraloop. In any of the foregoing or related aspects, L is a tetraloop. In some aspects, the tetraloop comprises the sequence 5′-GAAA-3′. In any of the foregoing or related aspects, S1 and S2 are 1-10 nucleotides in length and have the same length. In some aspects, S1 and S2 are 1 nucleotide, 2 nucleotides, 3 nucleotides, 4 nucleotides, 5 nucleotides, 6 nucleotides, 7 nucleotides, 8 nucleotides, 9 nucleotides, or 10 nucleotides in length. In some aspects, S1 and S2 are 6 nucleotides in length. In any of the foregoing or related aspects, the stem-loop comprises the sequence 5′-GCAGCCGAAAGGCUGC-3′ (SEQ ID NO: 871).

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide comprising a nicked tetraloop structure. In some aspects, the RNAi oligonucleotide comprises a nick between the 3′ terminus of the sense strand and the 5′ terminus of the antisense strand. In some aspects, the antisense and sense strands are not covalently linked.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the antisense strand comprises a 3′ overhang of one or more nucleotides in length. In some aspects, the 3′ overhang comprises purine nucleotides. In some aspects, the 3′ overhang is 2 nucleotides in length. In some aspects, the 3′ overhang is selected from AA, GG, AG and GA. In some aspects, the 3′ overhang is GG or AA. In some aspects, the 3′ overhang is GG.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide comprising at least one modified nucleotide. In some aspects, the modified nucleotide comprises a 2′-modification. In some aspects, the 2′-modification is a modification selected from 2′-aminoethyl, 2′-fluoro, 2′-O-methyl, 2′-O-methoxyethyl, and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid. In some aspects, the 2′-modification is 2′-fluoro. In some aspects, the 2′-modification is 2′-O-methyl. In some aspects, the 2′-modification is 2′-fluoro and 2′-O-methyl.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide comprising at least one modified nucleotide, wherein about 10-15%, 10%, 11%, 12%, 13%, 14% or 15% of the nucleotides of the sense strand comprise a 2′-fluoro modification. In some aspects, about 25-35%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34% or 35% of the nucleotides of the antisense strand comprise a 2′-fluoro modification. In some aspects, about 25-35%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34% or 35% of the nucleotides of the oligonucleotide comprise a 2′-fluoro modification. In some aspects, the sense strand comprises 36 nucleotides with positions 1-36 from 5′ to 3′, wherein positions 8-11 comprise a 2′-fluoro modification. In some aspects, the antisense strand comprises 22 nucleotides with positions 1-22 from 5′ to 3′, wherein positions 2, 3, 4, 5, 7, 10 and 14 comprise a 2′-fluoro modification. In some aspects, the remaining nucleotides of the sense and/or antisense strand comprise a 2′-O-methyl modification.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein all of the nucleotides are modified. In some aspects, positions 8, 9, 10 and 11 (from 5′ to 3′) of the sense strand are modified. In some aspects, positions 3, 8, 9, 10, 12, 13 and 17 (from 5′ to 3′) of the sense strand are modified. In some aspects, positions 2, 3, 4, 5, 7, 10 and 14 (from 5′ to 3′) of the antisense strand are modified. In some aspects, positions 2-5, 7, 8, 10, 14, 16 and 19 (from 5′ to 3′) of the antisense strand are modified. In some aspects, positions 8, 9, 10 and 11 (from 5′ to 3′) of the sense strand and positions 2, 3, 4, 5, 7, 10 and 14 (from 5′ to 3′) of the antisense strand are modified. In some aspects, positions 3, 8, 9, 10, 12, 13 and 17 (from 5′ to 3′) of the sense strand and positions 2-5, 7, 8, 10, 14, 16 and 19 (from 5′ to 3′) of the antisense strand are modified. In some aspects, the modification is a 2′-fluoro modification.

In any of the foregoing or related aspects, the oligonucleotide comprises at least one modified internucleotide linkage. In some aspects, the at least one modified internucleotide linkage is a phosphorothioate linkage. In some aspects, the antisense strand comprises a phosphorothioate linkage (i) between positions 1 and 2, and between positions 2 and 3; or (ii) between positions 1 and 2, between positions 2 and 3, and between positions 3 and 4, wherein positions are numbered 1-4 from 5′ to 3′. In some aspects, the antisense strand is 22 nucleotides in length and comprises a phosphorothioate linkage between positions 20 and 21 and between positions 21 and 22, wherein positions are numbered 1-22 from 5′ to 3′.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the 4′-carbon of the sugar of the 5′-nucleotide of the antisense strand comprises a phosphate analog. In some aspects, the phosphate analog is oxymethylphosphonate, vinylphosphonate or malonyl phosphonate, optionally wherein the phosphate analog is a 4′-phosphate analog comprising 5′-methoxyphosphonate-4′-oxy.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide comprising an antisense strand comprising a phosphorylated nucleotide at the 5′ terminus, wherein the phosphorylated nucleotide is selected from uridine and adenosine. In some aspects, the phosphorylated nucleotide is uridine.

In any of the foregoing or related aspects, the oligonucleotide reduces or inhibits KHK expression in vivo. In any of the foregoing or related aspects, the oligonucleotide is a Dicer substrate. In some aspects, the oligonucleotide is a Dicer substrate that, upon endogenous Dicer processing, yields double-stranded nucleic acids of 19-23 nucleotides in length capable of reducing KHK expression in a mammalian cell.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein at least one nucleotide of the oligonucleotide is conjugated to one or more targeting ligands. In some aspects, each targeting ligand comprises a carbohydrate, amino sugar, cholesterol, polypeptide, or lipid. In some aspects, the stem loop comprises one or more targeting ligands conjugated to one or more nucleotides of the stem loop. In some aspects, one or more targeting ligands is conjugated to one or more nucleotides of the loop. In some aspects, the loop comprises 4 nucleotides numbered 1-4 from 5′ to 3′, wherein nucleotides at positions 2, 3 and 4 each comprise one or more targeting ligands, wherein the targeting ligands are the same or different.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein each targeting ligand comprises a N-acetylgalactosamine (GalNAc) moiety. In some aspects, the GalNAc moiety is a monovalent GalNAc moiety, a bivalent GalNAc moiety, a trivalent GalNAc moiety or a tetravalent GalNAc moiety. In some aspects, up to 4 nucleotides of L of the stem-loop are each conjugated to a monovalent GalNAc moiety.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide comprising an antisense strand comprising a region of complementarity, wherein the region of complementarity is fully complementary to the KHK mRNA target sequence at nucleotide positions 2-8 of the antisense strand, wherein nucleotide positions are numbered 5′ to 3′. In some aspects, the region of complementarity is fully complementary to the KHK mRNA target sequence at nucleotide positions 2-11 of the antisense strand, wherein nucleotide positions are numbered 5′ to 3′.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs: 872-878 and 886-911.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the antisense strand comprises a nucleotide sequence of any one of

SEQ ID NOs: 879-884 and 912-938.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the antisense strand comprises a nucleotide sequence selected from SEQ ID NOs: 913, 917, 918, 920, 923 and 936. In some aspects, the sense strand comprises a nucleotide sequence selected from SEQ ID NOs: 942-947. In some aspects, the sense strand comprises a nucleotide sequence selected from SEQ ID NOs: 887, 891, 892, 894, 897 and 909.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 886 and 912, respectively;
  • (b) SEQ ID NOs: 887 and 913, respectively;
  • (c) SEQ ID NOs: 910 and 937, respectively;
  • (d) SEQ ID NOs: 888 and 914, respectively;
  • (e) SEQ ID NOs: 889 and 915, respectively;
  • (f) SEQ ID NOs: 890 and 916, respectively;
  • (g) SEQ ID NOs: 891 and 917, respectively;
  • (h) SEQ ID NOs: 877 and 884, respectively;
  • (i) SEQ ID NOs: 878 and 930, respectively;
  • (j) SEQ ID NOs: 876 and 883, respectively;
  • (k) SEQ ID NOs: 875 and 882, respectively;
  • (l) SEQ ID NOs: 892 and 918, respectively;
  • (m) SEQ ID NOs: 893 and 919, respectively;
  • (n) SEQ ID NOs: 894 and 920, respectively;
  • (o) SEQ ID NOs: 904 and 931, respectively;
  • (p) SEQ ID NOs: 895 and 921, respectively;
  • (q) SEQ ID NOs: 905 and 932, respectively;
  • (r) SEQ ID NOs: 896 and 922, respectively;
  • (s) SEQ ID NOs: 911 and 938, respectively;
  • (t) SEQ ID NOs: 906 and 933, respectively;
  • (u) SEQ ID NOs: 897 and 923, respectively;
  • (v) SEQ ID NOs: 907 and 934, respectively;
  • (w) SEQ ID NOs: 908 and 935, respectively;
  • (x) SEQ ID NOs: 903 and 929, respectively;
  • (y) SEQ ID NOs: 901 and 927, respectively;
  • (z) SEQ ID NOs: 874 and 881, respectively;
  • (aa) SEQ ID NOs: 902 and 928, respectively;
  • (bb) SEQ ID NOs: 873 and 880, respectively;
  • (cc) SEQ ID NOs: 872 and 879, respectively;
  • (dd) SEQ ID NOs: 898 and 924, respectively;
  • (ee) SEQ ID NOs: 899 and 925, respectively;
  • (gg) SEQ ID NOs: 900 and 926, respectively; and
  • (hh) SEQ ID NOs: 909 and 936, respectively.

In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 887 and 913, respectively;
  • (b) SEQ ID NOs: 891 and 917, respectively;
  • (c) SEQ ID NOs: 892 and 918, respectively;
  • (d) SEQ ID NOs: 894 and 920, respectively;
  • (e) SEQ ID NOs: 897 and 923, respectively; and
  • (f) SEQ ID NOs: 909 and 936, respectively.

In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 909 and 936, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 894 and 920, respectively. In yet other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 897 and 923, respectively. In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 892 and 918, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 891 and 917, respectively. In yet further aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 887 and 913, respectively.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the oligonucleotide as described herein achieves at least 50% knockdown of KHK mRNA. In some aspects, an oligonucleotide described herein achieves at least 50% knockdown of KHK mRNA in vitro. In some aspects, an oligonucleotide described herein achieves at least 50% knockdown of KHK mRNA in vivo. In some aspects, an oligonucleotide described herein achieves at least 50% knockdown of KHK mRNA in vitro and in vivo. In some aspects, an oligonucleotide described herein that achieves at least 50% knockdown of KHK mRNA comprises a sense strand and an antisense strand, wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 886 and 912, respectively;
  • (b) SEQ ID NOs: 887 and 913, respectively;
  • (c) SEQ ID NOs: 910 and 937, respectively;
  • (d) SEQ ID NOs: 890 and 916, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively;
  • (f) SEQ ID NOs: 892 and 918, respectively;
  • (g) SEQ ID NOs: 893 and 919, respectively;
  • (h) SEQ ID NOs: 894 and 920, respectively;
  • (i) SEQ ID NOs: 911 and 938, respectively;
  • (j) SEQ ID NOs: 899 and 925, respectively;
  • (k) SEQ ID NOs: 900 and 926, respectively;
  • (l) SEQ ID NOs: 909 and 936, respectively; and
  • (m) SEQ ID NOs: 897 and 923, respectively.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the sense strand and the antisense strand are modified, wherein the antisense strand and the sense strand comprise one or more 2′-fluoro and 2′-O-methyl modified nucleotides and at least one phosphorothioate linkage, wherein the 4′-carbon of the sugar of the 5′-nucleotide of the antisense strand comprises a phosphate analog.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs: 774-804.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the antisense strand comprises a nucleotide sequence of any one of

SEQ ID NOs: 819-849.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 774 and 819, respectively;
  • (b) SEQ ID NOs: 775 and 820, respectively;
  • (c) SEQ ID NOs: 776 and 821, respectively;
  • (d) SEQ ID NOs: 777 and 822, respectively;
  • (e) SEQ ID NOs: 778 and 823, respectively;
  • (f) SEQ ID NOs: 779 and 824, respectively;
  • (g) SEQ ID NOs: 780 and 825, respectively;
  • (h) SEQ ID NOs: 781 and 826, respectively;
  • (i) SEQ ID NOs: 782 and 827, respectively;
  • (j) SEQ ID NOs: 783 and 828, respectively;
  • (k) SEQ ID NOs: 784 and 829, respectively;
  • (l) SEQ ID NOs: 785 and 830, respectively;
  • (m) SEQ ID NOs: 786 and 831, respectively;
  • (n) SEQ ID NOs: 787 and 832, respectively;
  • (o) SEQ ID NOs: 788 and 833, respectively;
  • (p) SEQ ID NOs: 789 and 834, respectively;
  • (q) SEQ ID NOs: 790 and 835, respectively;
  • (r) SEQ ID NOs: 791 and 836, respectively;
  • (s) SEQ ID NOs: 792 and 837, respectively;
  • (t) SEQ ID NOs: 793 and 838, respectively;
  • (u) SEQ ID NOs: 794 and 839, respectively;
  • (v) SEQ ID NOs: 795 and 840, respectively;
  • (w) SEQ ID NOs: 796 and 841, respectively;
  • (x) SEQ ID NOs: 797 and 842, respectively;
  • (y) SEQ ID NOs: 798 and 843, respectively;
  • (z) SEQ ID NOs: 799 and 844, respectively;
  • (aa) SEQ ID NOs: 800 and 845, respectively;
  • (bb) SEQ ID NOs: 801 and 846, respectively;
  • (cc) SEQ ID NOs: 802 and 847, respectively;
  • (dd) SEQ ID NOs: 803 and 848, respectively; and
  • (ee) SEQ ID NOs: 804 and 849, respectively.

In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 775 and 820, respectively;
  • (b) SEQ ID NOs: 779 and 824, respectively;
  • (c) SEQ ID NOs: 780 and 825, respectively;
  • (d) SEQ ID NOs: 782 and 827, respectively;
  • (e) SEQ ID NOs: 785 and 830, respectively; and
  • (f) SEQ ID NOs: 804 and 849, respectively.

In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 804 and 849, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 782 and 827, respectively. In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 775 and 820, respectively. In yet other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 779 and 824, respectively. In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 780 and 825, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 785 and 830, respectively.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs: 805-818.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the antisense strand comprises a nucleotide sequence of any one of

SEQ ID NOs: 850-863.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 805 and 850, respectively;
  • (b) SEQ ID NOs: 806 and 851, respectively;
  • (c) SEQ ID NOs: 807 and 852, respectively;
  • (d) SEQ ID NOs: 808 and 853, respectively;
  • (e) SEQ ID NOs: 809 and 854, respectively;
  • (f) SEQ ID NOs: 810 and 855, respectively;
  • (g) SEQ ID NOs: 811 and 856, respectively;
  • (h) SEQ ID NOs: 812 and 857, respectively;
  • (i) SEQ ID NOs: 813 and 858, respectively;
  • (j) SEQ ID NOs: 814 and 859, respectively;
  • (k) SEQ ID NOs: 815 and 860, respectively;
  • (l) SEQ ID NOs: 816 and 861, respectively;
  • (m) SEQ ID NOs: 817 and 862, respectively and;
  • (n) SEQ ID NOs: 818 and 863, respectively.

In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences

set forth in SEQ ID NOs: 805 and 850, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 809 and 854, respectively. In yet other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 810 and 855, respectively. In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 812 and 857, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 815 and 860, respectively. In yet other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 818 and 863, respectively.

In some aspects, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mG-S-mA-mA-mG-mA-mG-mA-fA-fG-fC-fA-mG-mA-mU-mC-mC-mU-mG-mU-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 775), and wherein the antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fA-S-fC-fA-fG-mG-fA-mU-mC-fU-mG-mC-mU-fU-mC-mU-mC-mU-mU-mC-S-mG-S-mG-3′ (SEQ ID NO: 820), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.

In some aspects, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mC-S-mA-mG-mA-mU-mG-mU-fG-fU-fC-fU-mG-mC-mU-mA-mC-mA-mG-mA-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 779), and wherein the antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fU-S-fC-S-fU-fG-mU-fA-mG-mC-fA-mG-mA-mC-fA-mC-mA-mU-mC-mU-mG-S-mG-S-mG-3′ (SEQ ID NO: 824), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.

In some aspects, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mG-S-mA-mC-mU-mU-mU-mG-fA-fG-fA-fA-mG-mG-mU-mU-mG-mA-mU-mC-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 780), and wherein the antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fG-S-fA-S-fU-fC-mA-fA-mC-mC-fU-mU-mC-mU-fC-mA-mA-mA-mG-mU-mC-S-mG-S-mG-3′ (SEQ ID NO: 825), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.

In some aspects, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mU-S-mG-mU-mU-mU-mG-mU-fC-fA-fG-fC-mA-mA-mA-mG-mA-mU-mG-mU-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 785), and wherein the antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fA-S-fC-fA-fU-mC-fU-mU-mU-fG-mC-mU-mG-fA-mC-mA-mA-mA-mC-mA-S-mG-S-mG-3′ (SEQ ID NO: 830), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.

In some aspects, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mG-S-mC-mA-mG-mG-mA-mA-fG-fC-fA-fC-mU-mG-mA-mG-mA-mU-mU-mC-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 804), and wherein the antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fG-S-fA-S-fA-fU-mC-fU-mC-mA-fG-mU-mG-mC-fU-mU-mC-mC-mU-mG-mC-S-mG-S-mG-3′ (SEQ ID NO: 849), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.

In some aspects, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNA comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mU-S-mU-mU-mG-mA-mG-mA-fA-fG-fG-fU-mU-mG-mA-mU-mC-mU-mG-mA-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 782), and wherein the antisense strand comprises the sequence and all of the modifications of 5′ [MePhosphonate-4O-mU]-S-fU-S-fC-S-fA-fG-mA-fU-mC-mA-fA-mC-mC-mU-fU-mC-mU-mC-mA-mA-mA-S-mG-S-mG-3′ (SEQ ID NO: 827), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.

In yet other aspects, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNA comprises a sense strand comprising SEQ ID NO: 775 and an antisense strand comprising SEQ ID NO: 820, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNA is in the form of a conjugate having the structure depicted in FIG. 10A continuing to FIG. 10B, or pharmaceutically acceptable salts thereof.

In another aspect, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNA comprises a sense strand comprising SEQ ID NO: 779 and an antisense strand comprising SEQ ID NO: 824, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNA is in the form of a conjugate having the structure depicted in FIG. 11A continuing to FIG. 11B, or pharmaceutically acceptable salts thereof.

In another aspect, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNA comprises a sense strand comprising SEQ ID NO: 780 and an antisense strand comprising SEQ ID NO: 825, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNA is in the form of a conjugate having the structure depicted in FIG. 12A continuing to FIG. 12B, or pharmaceutically acceptable salts thereof.

In another aspect, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNA comprises a sense strand comprising SEQ ID NO: 782 and an antisense strand comprising SEQ ID NO: 827, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNA is in the form of a conjugate having the structure depicted in FIG. 13A continuing to FIG. 13B, or pharmaceutically acceptable salts thereof.

In another aspect, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNA comprises a sense strand comprising SEQ ID NO: 785 and an antisense strand comprising SEQ ID NO: 830, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNA is in the form of a conjugate having the structure depicted in FIG. 14A continuing to FIG. 14B, or pharmaceutically acceptable salts thereof.

In another aspect, the present disclosure provides a double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNA comprises a sense strand comprising SEQ ID NO: 804 and an antisense strand comprising SEQ ID NO: 849, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNA is in the form of a conjugate having the structure depicted in FIG. 15A continuing to FIG. 15B, or pharmaceutically acceptable salts thereof.

In some aspects, the present disclosure provides a method for treating a subject having a disease, disorder or condition associated with KHK expression, the method comprising administering to the subject a therapeutically effective amount of any RNAi oligonucleotide or pharmaceutical composition described herein, thereby treating the subject.

In some aspects, the disclosure provides a pharmaceutically acceptable salt of any of the oligonucleotides described herein. In some aspects, the present disclosure provides a pharmaceutical composition comprising any RNAi oligonucleotide described herein, and a pharmaceutically acceptable carrier, salt, delivery agent or excipient. In some aspects, the present disclosure provides a pharmaceutical composition comprising any RNAi oligonucleotide described herein, and a pharmaceutically acceptable diluent, solvent, carrier, salt and/or adjuvant. Likewise, the oligonucleotides herein may be provided in the form of their free acids.

In some aspects, the disclosure provides a method for modulating KHK expression in a target cell expressing KHK, the method comprising administering an RNAi oligonucleotide or pharmaceutical composition described herein in an effective amount to the target cell.

In some aspects, the present disclosure provides a method of delivering an oligonucleotide to a subject, the method comprising administering a pharmaceutical composition described herein.

In some aspects, the present disclosure provides a method for reducing KHK expression in a cell, a population of cells or a subject, the method comprising the step of:

• • i. contacting the cell or the population of cells with any RNAi oligonucleotide or pharmaceutical composition described herein; or
  • ii. administering to the subject any RNAi oligonucleotide or pharmaceutical composition described herein.

In any of the foregoing or related aspects, the method of reducing KHK expression comprises reducing an amount or level of KHK mRNA, an amount or level of KHK protein, or both.

In any of the foregoing or related aspects, the subject has a disease, disorder or condition associated with KHK expression. In some aspects, the disease, disorder, or condition associated with KHK expression is non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

In any of the foregoing or related aspects, the RNAi oligonucleotide, or pharmaceutical composition, is administered in combination with a second composition or therapeutic agent.

In some aspects, the present disclosure provides a method for treating a subject having a disease, disorder or condition associated with KHK expression, the method comprising administering to the subject a therapeutically effective amount of an RNAi oligonucleotide comprising a sense strand and an antisense strand, or a pharmaceutically acceptable salt thereof, wherein the sense strand and antisense strand comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 886 and 912, respectively;
  • (b) SEQ ID NOs: 887 and 913, respectively;
  • (c) SEQ ID NOs: 910 and 937, respectively;
  • (d) SEQ ID NOs: 888 and 914, respectively;
  • (e) SEQ ID NOs: 889 and 915, respectively;
  • (f) SEQ ID NOs: 890 and 916, respectively;
  • (g) SEQ ID NOs: 891 and 917, respectively;
  • (h) SEQ ID NOs: 877 and 884, respectively;
  • (i) SEQ ID NOs: 878 and 930, respectively;
  • (j) SEQ ID NOs: 876 and 883, respectively;
  • (k) SEQ ID NOs: 875 and 882, respectively;
  • (l) SEQ ID NOs: 892 and 918, respectively;
  • (m) SEQ ID NOs: 893 and 919, respectively;
  • (n) SEQ ID NOs: 894 and 920, respectively;
  • (o) SEQ ID NOs: 904 and 931, respectively;
  • (p) SEQ ID NOs: 895 and 921, respectively;
  • (q) SEQ ID NOs: 905 and 932, respectively;
  • (r) SEQ ID NOs: 896 and 922, respectively;
  • (s) SEQ ID NOs: 911 and 938, respectively;
  • (t) SEQ ID NOs: 906 and 933, respectively;
  • (u) SEQ ID NOs: 897 and 923, respectively;
  • (v) SEQ ID NOs: 907 and 934, respectively;
  • (w) SEQ ID NOs: 908 and 935, respectively;
  • (x) SEQ ID NOs: 903 and 929, respectively;
  • (y) SEQ ID NOs: 901 and 927, respectively;
  • (z) SEQ ID NOs: 874 and 881, respectively;
  • (aa) SEQ ID NOs: 902 and 928, respectively;
  • (bb) SEQ ID NOs: 873 and 880, respectively;
  • (cc) SEQ ID NOs: 872 and 879, respectively;
  • (dd) SEQ ID NOs: 898 and 924, respectively;
  • (ee) SEQ ID NOs: 899 and 925, respectively
  • (ff) SEQ ID NOs: 900 and 926, respectively; and
  • (gg) SEQ ID NOs: 909 and 936, respectively.

In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 887 and 913, respectively;
  • (b) SEQ ID NOs: 891 and 917, respectively;
  • (c) SEQ ID NOs: 892 and 918, respectively;
  • (d) SEQ ID NOs: 894 and 920, respectively;
  • (e) SEQ ID NOs: 897 and 923, respectively; and
  • (f) SEQ ID NOs: 909 and 936, respectively.

In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 887 and 913, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 891 and 917, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 892 and 918, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 894 and 920, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 897 and 923, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 909 and 936, respectively.

In some aspects, the present disclosure provides a method for treating a subject having a disease, disorder or condition associated with KHK expression, the method comprising administering to the subject a therapeutically effective amount of an RNAi oligonucleotide comprising a sense strand and an antisense strand, or a pharmaceutically acceptable salt thereof, wherein the sense strand and antisense strands are selected from the group consisting of:

• • (a) SEQ ID NOs: 774 and 819, respectively;
  • (b) SEQ ID NOs: 775 and 820, respectively;
  • (c) SEQ ID NOs: 776 and 821, respectively;
  • (d) SEQ ID NOs: 777 and 822, respectively;
  • (e) SEQ ID NOs: 778 and 823, respectively;
  • (f) SEQ ID NOs: 779 and 824, respectively;
  • (g) SEQ ID NOs: 780 and 825, respectively;
  • (h) SEQ ID NOs: 781 and 826, respectively;
  • (i) SEQ ID NOs: 782 and 827, respectively;
  • (j) SEQ ID NOs: 783 and 828, respectively;
  • (k) SEQ ID NOs: 784 and 829, respectively;
  • (l) SEQ ID NOs: 785 and 830, respectively;
  • (m) SEQ ID NOs: 786 and 831, respectively;
  • (n) SEQ ID NOs: 787 and 832, respectively;
  • (o) SEQ ID NOs: 788 and 833, respectively;
  • (p) SEQ ID NOs: 789 and 834, respectively;
  • (q) SEQ ID NOs: 790 and 835, respectively;
  • (r) SEQ ID NOs: 791 and 836, respectively;
  • (s) SEQ ID NOs: 792 and 837, respectively;
  • (t) SEQ ID NOs: 793 and 838, respectively;
  • (u) SEQ ID NOs: 794 and 839, respectively;
  • (v) SEQ ID NOs: 795 and 840, respectively;
  • (w) SEQ ID NOs: 796 and 841, respectively;
  • (x) SEQ ID NOs: 797 and 842, respectively;
  • (y) SEQ ID NOs: 798 and 843, respectively;
  • (z) SEQ ID NOs: 799 and 844, respectively;
  • (aa) SEQ ID NOs: 800 and 845, respectively;
  • (bb) SEQ ID NOs: 801 and 846, respectively;
  • (cc) SEQ ID NOs: 802 and 847, respectively;
  • (dd) SEQ ID NOs: 803 and 848, respectively; and
  • (ee) SEQ ID NOs: 804 and 849, respectively.

In any of the foregoing or related aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 775 and 820, respectively;
  • (b) SEQ ID NOs: 779 and 824, respectively;
  • (c) SEQ ID NOs: 780 and 825, respectively;
  • (d) SEQ ID NOs: 782 and 827, respectively;
  • (e) SEQ ID NOs: 785 and 830, respectively; and
  • (f) SEQ ID NOs: 804 and 849, respectively.

In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 775 and 820, respectively. In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 779 and 824, respectively. In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 780 and 825, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 782 and 827, respectively. In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 785 and 830, respectively. In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 804 and 849, respectively.

In some aspects, the present disclosure provides a method for treating a subject having a disease, disorder or condition associated with KHK expression, the method comprising administering to the subject a therapeutically effective amount of an RNAi oligonucleotide comprising a sense strand and an antisense strand, or a pharmaceutically acceptable salt thereof, wherein the sense strand and antisense strands are selected from the group consisting of:

• • (a) SEQ ID NOs: 805 and 850, respectively;
  • (b) SEQ ID NOs: 806 and 851, respectively;
  • (c) SEQ ID NOs: 807 and 852, respectively;
  • (d) SEQ ID NOs: 808 and 853, respectively;
  • (e) SEQ ID NOs: 809 and 854, respectively;
  • (f) SEQ ID NOs: 810 and 855, respectively;
  • (g) SEQ ID NOs: 811 and 856, respectively;
  • (h) SEQ ID NOs: 812 and 857, respectively;
  • (i) SEQ ID NOs: 813 and 858, respectively;
  • (j) SEQ ID NOs: 814 and 859, respectively;
  • (k) SEQ ID NOs: 815 and 860, respectively;
  • (l) SEQ ID NOs: 816 and 861, respectively;
  • (m) SEQ ID NOs: 817 and 862, respectively and;
  • (n) SEQ ID NOs: 818 and 863, respectively.

In some aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 805 and 850, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 809 and 854, respectively. In yet other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 810 and 855, respectively. In further aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 812 and 857, respectively. In other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 815 and 860, respectively. In yet other aspects, the disclosure provides an RNAi oligonucleotide wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 818 and 863, respectively.

In any of the foregoing or related aspects, the disease, disorder, or condition associated with KHK expression is non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

In any of the foregoing or related aspects, a RNAi oligonucleotide described herein is administered at a concentration of 0.01 mg/kg-5 mg/kg.

In some aspects, the disclosure provides use of any RNAi oligonucleotide or pharmaceutical composition described herein, in the manufacture of a medicament for the treatment of a disease, disorder or condition associated with KHK expression, optionally for the treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

In some aspects, the disclosure provides any RNAi oligonucleotide or pharmaceutical composition described herein, for use, or adaptable for use, in the treatment of a disease, disorder or condition associated with KHK expression, optionally for the treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

In some aspects, the disclosure provides a kit comprising any RNAi oligonucleotide described herein, an optional pharmaceutically acceptable carrier, and a package insert comprising instructions for administration to a subject having a disease, disorder or condition associated with KHK expression.

In some aspects, the disease, disorder, or condition associated with KHK expression is non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

In some aspects, the disclosure provides an oligonucleotide for reducing KHK expression, the oligonucleotide comprising a nucleotide sequence of 15-50 nucleotides in length, wherein the nucleotide sequence comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is at least 15 contiguous nucleotides in length, or a pharmaceutically acceptable salt thereof. In some aspects, the oligonucleotide is single stranded. In some aspects, the oligonucleotide is an antisense oligonucleotide. In some aspects, the nucleotide sequence is 15-30 nucleotides in length. In some aspects, the nucleotide sequence is 20-25 nucleotides in length. In some aspects, the nucleotide sequence is 22 nucleotides in length. In some aspects, the region of complementarity is 19 contiguous nucleotides in length. In some aspects, the region of complementarity is 20 contiguous nucleotides in length. In some aspects, the nucleotide sequence comprises at least one modification. In some aspects, the nucleotide sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 879-885 and 912-938. In some aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 920. In other aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 923. In yet other aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 918. In further aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 917. In yet further aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 913. In yet further aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 936. In some aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 894. In other aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 897. In yet other aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 892. In further aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 891. In yet further aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 887. In yet further aspects, the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 909.

In some aspects, the disclosure provides a cell comprising an oligonucleotide described herein.

In some aspects, the disclosure provides a pharmaceutical composition comprising an oligonucleotide disclosed herein, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, delivery agent or excipient.

In some aspects, the disclosure provides a method for treating a subject having a disease, disorder or condition associated with KHK expression, the method comprising administering to the subject a therapeutically effective amount of an oligonucleotide or pharmaceutical composition described herein.

In some aspects, the disclosure provides a method of delivering an oligonucleotide to a subject, the method comprising administering a pharmaceutical composition described herein to the subject.

In some aspects, the disclosure provides a method for reducing KHK expression in a cell, a population of cells or a subject, the method comprising the step of:

i. contacting the cell or the population of cells with an oligonucleotide or a pharmaceutical composition described herein; orii. administering to the subject an oligonucleotide or a pharmaceutical composition described herein. In some aspects, reducing KHK expression comprises reducing an amount or level of KHK mRNA, an amount or level of KHK protein, or both.

In any of the foregoing or related aspects, the subject has a disease, disorder or condition associated with KHK expression. In some aspects, the disease, disorder, or condition associated with KHK expression is non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

In any of the foregoing or related aspects, the oligonucleotide, or pharmaceutical composition, is administered in combination with a second composition or therapeutic agent.

In some aspects, the disclosure provides use of an oligonucleotide or pharmaceutical composition described herein, in the manufacture of a medicament for the treatment of a disease, disorder or condition associated with KHK expression, optionally for the treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). In other aspects, the disclosure provides an oligonucleotide or pharmaceutical composition described herein for use, or adaptable for use, in the treatment of a disease, disorder or condition associated with KHK expression, optionally for the treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

In some aspects, the disclosure provides a kit comprising an oligonucleotide described herein, an optional pharmaceutically acceptable carrier, and a package insert comprising instructions for administration to a subject having a disease, disorder or condition associated with KHK expression.

In any of the foregoing or related aspects, the disease, disorder, or condition associated with KHK expression is non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

In some aspects, the disclosure provides a double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of KHK, wherein the dsRNA agent comprises a sense strand and an antisense strand forming a duplex region, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from a nucleotide sequence selected from SEQ ID NOs: 4-387, and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from a nucleotide sequence selected from SEQ ID NOs: 388-771.

In some aspects, the disclosure provides a double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of KHK, wherein the dsRNA agent comprises a sense strand and an antisense strand forming a duplex region, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from a nucleotide sequence selected from SEQ ID NOs: 872-878 and 886-911, and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from a nucleotide sequence selected from SEQ ID NOs: 879-885 and 912-938.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 provides a graph depicting the percent (%) mRNA remaining in Hep3B cells (expressing endogenous human KHK) after 24-hour treatment with 1 nM of DsiRNA targeting various regions of the KHK gene. 384 DsiRNAs were designed and screened. Three primer pairs were used that recognized the KHK-A isoform (KHK-F763, NM_000221.2), KHK-C (KHK-825, NM_006488.3) and KHK-All (both isoforms) (KHK-F495, KHK-F1026, NM_006488.3). Expression was normalized between samples using HPRT and SFRS9 housekeeping genes.

FIG. 2A and FIG. 2B provide schematics of the Low-2′-Fluoro modification pattern (Low-2′-Fluoro (3PS) and Low-2′-Fluoro (2PS), respectively) applied to KHK mRNA targeting sequences to generate GalNAc-KHK constructs. The sense strand includes a tetraloop structure of nucleotides 26-31 of the 36-nucleotide strand. The anti-sense strand is complementary and includes a 2-nucleotide overhang.

FIG. 3 provides a graph depicting the percent (%) remaining KHK mRNA in KHK-A and KHK-C HDI (hydrodynamic injection) mice treated with human/non-human primate (NHP)-conserved GalNAc-KHK constructs. 3 days after subcutaneous dosing of 2 mg/kg of

GalNAc-KHK constructs formulated in PBS, plasmids encoding either KHK-A and KHK-C were injected into mice via HDI and the percent (%) of KHK mRNA was measured 1 day later in liver samples relative to mice treated with PBS. mRNA was measured from livers using primers recognizing KHK-All (up-right triangle), KHK-C (upside-down triangle), and KHK-A (hexagon). The notation “Hs, 1 mm Mf” represents a human specific sequence that is one base mismatch different from monkey sequence.

FIG. 4A provides a schematic of the Med-2′-Fluoro modification pattern applied to KHK targeting sequence to generate GalNAc-KHK constructs. The sense strand includes a tetraloop structure of nucleotides 26-31 of the 36-nucleotide strand. The anti-sense strand is complementary and includes a 2-nucleotide overhang.

FIGS. 4B-4C provide graphs depicting the percent (%) KHK mRNA remaining after treating mice with GalNAc-KHK constructs having the Med-2′-Fluoro modification pattern. 3 days after subcutaneous dosing of 2 mg/kg of GalNAc-KHK constructs formulated in PBS, plasmid encoding KHK-C was injected into mice via HDI and the percent (%) of KHK mRNA was measured 1 day later in liver samples relative to mice treated with PBS. mRNA was measured using primers identifying both KHK-A and KHK-C isoform s (i.e., KHK-All) (FIG. 4B) and primers identifying only the KHK-C isoform (FIG. 4C). Multiple GalNAc-KHK-constructs were combined in a “mixed” group at 2 mg/kg for a total 10 mg/kg treatment (KHK-0861, -0865, -0882, -0883, -0885) as a positive knock-down control. The notation “Hs, 1 mm Mf” and the like represents a human specific sequence that is one base mismatch different from monkey sequence.

FIG. 4D provides a graph depicting the percent (%) KHK mRNA remaining after treating mice with different GalNAc-KHK constructs having the Med-2′-Fluoro modification pattern. 3 days after subcutaneous dosing of 2 mg/kg of GalNAc-KHK constructs formulated in PBS, plasmids encoding KHK-C were injected into mice via HDI and the percent (%) of KHK mRNA was measured 1 day later in liver samples relative to mice treated with PBS. mRNA was measured using primers (MmKHK-ALL-5-6, Forward: GCTCTTCCAGTTGTTTAGCTATGGT (SEQ ID NO: 939), Reverse: CAGGTGCTTGGCCACATCTT (SEQ ID NO:940), Probe: AGGTGGTGTTTGTCAGC (SEQ ID NO: 941)) identifying only mouse KHK. Remaining mRNA was normalized to a PBS control. Multiple GalNAc-KHK constructs were combined in a “mixed” group as a positive knock-down control.

FIG. 4E provides a graph depicting the difference in percent (%) KHK mRNA remaining after treatment with GalNAc-KHK constructs with different modification patterns (Low-2′F (FIG. 2A and FIG. 2B) and Med-2′F (FIG. 4A)). Remaining mRNA was normalized to a PBS control. Multiple GalNAc-KHK constructs were combined in a “mixed” group as a positive knock-down control.

FIG. 5 provides a graph depicting the difference in percent (%) KHK mRNA remaining after treating mice with GalNAc-KHK constructs. 3 days after subcutaneous dosing of 2 mg/kg of GalNAc-KHK constructs formulated in PBS, plasmid encoding KHK-C(NM_006488) (pCMV6-KHK-C, Cat #: RC223488, OriGene) was injected into mice via HDI and the percent (%) of KHK mRNA remaining was measured 1 day later in liver samples relative to mice treated with PBS. Results include mRNA measured from primers for KHK-All (up-right triangle), and KHK-C (upside-down triangle). Grey arrow shows 30 mg/kg treatment of KHK-885 has more than 98% knockdown.

FIGS. 6A-6B provide graphs depicting the percent (%) KHK mRNA remaining after treating KHK-C plasmid HDI mice (as described in FIG. 5) with different GalNAc-KHK constructs. mRNA was measured using primers identifying both KHK-A and KHK-C isoforms (KHK-All; FIG. 6A) and primers identifying only the KHK-C isoform (FIG. 6B).

FIG. 6C provides a graph depicting the percent (%) KHK mRNA remaining in the liver after treating KHK-C plasmid HDI mice (as described in FIG. 5) with different GalNAc-KHK constructs. mRNA was measured using primers identifying only mouse KHK.

FIGS. 7A-7C provide graphs depicting the percent (%) KHK mRNA remaining in liver biopsies from non-human primates (NHP) 28 days (FIG. 7A), 56 days (FIG. 7B), and 84 days (FIG. 7C) after a single dose of specified GalNAc-constructs. NHP were subcutaneously injected with 6 mg/kg of GalNAc-KHK on Study Day 0. The percent indicated is the average reduction in KHK-mRNA compared to a PBS control.

FIG. 7D provides a line graph demonstrating the changes in KHK mRNA in liver biopsies taken at various time points from NHP (as treated in FIGS. 7A-7C) after a single dose of GalNAc-KHK constructs.

FIGS. 8A-8C provide graphs depicting the percent (%) KHK protein remaining in liver biopsies from non-human primates (NHP) 28 days (FIG. 8A), 56 days (FIG. 8B), and 84 days (FIG. 8C) after treatment. NHP were treated as in FIGS. 7A-7C. The percent indicated is the average reduction in KHK-protein compared to a PBS control.

FIG. 8D provides a line graph demonstrating the changes in KHK protein in liver biopsies taken at various time points from NHP (as treated in FIGS. 7A-7C) after a single dose of GalNAc-KHK constructs.

FIGS. 9A-9C provide correlation graphs demonstrating the relationship between remaining KHK mRNA expression and remaining KHK protein expression in liver biopsies from NHP treated with a single dose of GalNAc-KHK constructs. Correlation among all constructs is compared at days 28 (FIG. 9A), 56 (FIG. 9B), and 84 (FIG. 9C) after dosing. Individual dots represent individual biopsies.

FIG. 10A continuing to FIG. 10B depict a dsRNAi oligonucleotide of the invention comprising a sense strand comprising SEQ ID NO: 775 and an antisense strand comprising SEQ ID NO: 820, wherein said dsRNA is in the form of a conjugate.

FIG. 11A continuing to FIG. 11B depict a dsRNAi oligonucleotide of the invention comprising a sense strand comprising SEQ ID NO: 779 and an antisense strand comprising SEQ ID NO: 824, wherein said dsRNA is in the form of a conjugate.

FIG. 12A continuing to FIG. 12B depict a dsRNAi oligonucleotide of the invention comprising a sense strand comprising SEQ ID NO: 780 and an antisense strand comprising SEQ ID NO: 825, wherein said dsRNA is in the form of a conjugate.

FIG. 13A continuing to FIG. 13B depict a dsRNAi oligonucleotide of the invention comprising a sense strand comprising SEQ ID NO: 782 and an antisense strand comprising SEQ ID NO: 827, wherein said dsRNA is in the form of a conjugate.

FIG. 14A continuing to FIG. 14B depict a dsRNAi oligonucleotide of the invention comprising a sense strand comprising SEQ ID NO: 785 and an antisense strand comprising SEQ ID NO: 830, wherein said dsRNA is in the form of a conjugate.

FIG. 15A continuing to FIG. 15B depict a dsRNAi oligonucleotide of the invention comprising a sense strand comprising SEQ ID NO: 804 and an antisense strand comprising SEQ ID NO: 849, wherein said dsRNA is in the form of a conjugate.

DETAILED DESCRIPTION OF THE INVENTION

According to some aspects, the disclosure provides oligonucleotides that reduce KHK expression in the liver. In some embodiments, the oligonucleotides provided herein are useful to treat diseases associated with KHK expression in the liver. In some respects, the disclosure provides methods of treating a disease associated with KHK expression by reducing KHK gene expression in cells (e.g., cells of the liver).

Oligonucleotide Inhibitors of KHK Expression

Ketohexokinase (KHK) Target Sequences

In some embodiments, the disclosure provides an oligonucleotide which is targeted to a target sequence comprising a ketohexokinase (KHK) mRNA. In some embodiments, the oligonucleotide, or a portion, fragment, or strand thereof (e.g., an antisense strand or a guide strand of a dsRNA) binds or anneals to a target sequence comprising a KHK mRNA, thereby inhibiting KHK expression. In some embodiments, the oligonucleotide is targeted to a target sequence comprising a KHK-A isoform mRNA. In some embodiments, the oligonucleotide is targeted to a target sequence comprising a KHK-C isoform mRNA. In some embodiments, the oligonucleotide is targeted to a KHK target sequence for the purpose of inhibiting KHK expression in vivo. In some embodiments, the amount or extent of inhibition of KHK expression by an oligonucleotide targeted to a KHK target sequence correlates with the potency of the oligonucleotide. In some embodiments, the amount or extent of inhibition of KHK expression by an oligonucleotide targeted to a KHK target sequence correlates with the amount or extent of therapeutic benefit in a subject or patient having a disease, disorder or condition associated with the expression of KHK treated with the oligonucleotide.

Through examination of the nucleotide sequence of mRNAs encoding KHK, including mRNAs of multiple different species (e.g., human, cynomolgus monkey, mouse, and rat; see, e.g., Example 2) and as a result of in vitro and in vivo testing (see, e.g., Examples 2-6), it has been discovered that certain nucleotide sequences of KHK mRNA are more amenable than others to oligonucleotide-based inhibition and are thus useful as target sequences for the oligonucleotides herein. In some embodiments, a sense strand of an oligonucleotide (e.g., a dsRNA) described herein comprises a KHK target sequence. In some embodiments, a portion or region of the sense strand of a dsRNA described herein comprises a KHK target sequence. In some embodiments, a KHK target sequence comprises, or consists of, a sequence of any one of SEQ ID Nos: 4-387. In some embodiments, a KHK target sequence comprises, or consists of, nucleotides 1-19 of any one of SEQ ID Nos: 4-387. In some embodiments, a KHK target sequence comprises, or consists of, the sequence set forth in SEQ ID No: 39. In some embodiments, a KHK target sequence comprises, or consists of, nucleotides 1-19 of the sequence set forth in SEQ ID No: 39. In some embodiments, a KHK target sequence comprises, or consists of, the sequence set forth in SEQ ID No: 102. In some embodiments, a KHK target sequence comprises, or consists of, nucleotides 1-19 of the sequence set forth in SEQ ID No: 102. In some embodiments, a KHK target sequence comprises, or consists of, the sequence set forth in SEQ ID No: 104. In some embodiments, a KHK target sequence comprises, or consists of, nucleotides 1-19 of the sequence set forth in SEQ ID No: 104. In some embodiments, a KHK target sequence comprises, or consists of, the sequence set forth in SEQ ID No: 107. In some embodiments, a KHK target sequence comprises, or consists of, nucleotides 1-19 of the sequence set forth in SEQ ID No: 107. In some embodiments, a KHK target sequence comprises, or consists of, the sequence set forth in SEQ ID No: 191. In some embodiments, a KHK target sequence comprises, or consists of, nucleotides 1-19 of the sequence set forth in SEQ ID No: 191. In some embodiments, a KHK target sequence comprises, or consists of, the sequence set forth in SEQ ID No: 269. In some embodiments, a KHK target sequence comprises, or consists of, nucleotides 1-19 of the sequence set forth in SEQ ID No: 269.

KHK Targeting Sequences

In some embodiments, the oligonucleotides herein have regions of complementarity to KHK mRNA (e.g., within a target sequence of KHK mRNA) for purposes of targeting the mRNA in cells and inhibiting its expression. In some embodiments, the oligonucleotides herein comprise a KHK targeting sequence (e.g., an antisense strand or a guide strand of a dsRNA) having a region of complementarity that binds or anneals to a KHK target sequence by complementary (Watson-Crick) base pairing. The targeting sequence or region of complementarity is generally of a suitable length and base content to enable binding or annealing of the oligonucleotide (or a strand thereof) to a KHK mRNA for purposes of inhibiting its expression. In some embodiments, the targeting sequence or region of complementarity is at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21, at least about 22, at least about 23, at least about 24, at least about 25, at least about 26, at least about 27, at least about 28, at least about 29 or at least about 30 nucleotides in length. In some embodiments, the targeting sequence or region of complementarity is about 12 to about 30 (e.g., 12 to 30, 12 to 22, 15 to 25, 17 to 21, 18 to 27, 19 to 27, or 15 to 30) nucleotides in length. In some embodiments, the targeting sequence or region of complementarity is about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 nucleotides in length. In some embodiments, the targeting sequence or region of complementarity is 18 nucleotides in length. In some embodiments, the targeting sequence or region of complementarity is 19 nucleotides in length. In some embodiments, the targeting sequence or region of complementarity is 20 nucleotides in length. In some embodiments, the targeting sequence or region of complementarity is 21 nucleotides in length. In some embodiments, the targeting sequence or region of complementarity is 22 nucleotides in length. In some embodiments, the targeting sequence or region of complementarity is 23 nucleotides in length. In some embodiments, the targeting sequence or region of complementarity is 24 nucleotides in length. In some embodiments, an oligonucleotide comprises a target sequence or region of complementarity complementary to a sequence of any one of SEQ ID NOs: 4-387, and the targeting sequence or region of complementarity is 18 nucleotides in length. In some embodiments, an oligonucleotide comprises a target sequence or region of complementarity complementary to a sequence of any one of SEQ ID NOs: 4-387, and the targeting sequence or region of complementarity is 19 nucleotides in length. In some embodiments, an oligonucleotide comprises a target sequence or region of complementarity complementary to a sequence of any one of SEQ ID NOs: 4-387, and the targeting sequence or region of complementarity is 20 nucleotides in length. In some embodiments, an oligonucleotide comprises a target sequence or region of complementarity complementary to a sequence of any one of SEQ ID NOs: 4-387, and the targeting sequence or region of complementarity is 21 nucleotides in length. In some embodiments, an oligonucleotide comprises a target sequence or region of complementarity complementary to a sequence of any one of SEQ ID NOs: 4-387, and the targeting sequence or region of complementarity is 22 nucleotides in length. In some embodiments, an oligonucleotide comprises a target sequence or region of complementarity complementary to a sequence of any one of SEQ ID NOs: 4-387, and the targeting sequence or region of complementarity is 23 nucleotides in length. In some embodiments, an oligonucleotide comprises a target sequence or region of complementarity complementary to a sequence of any one of SEQ ID NOs: 4-387, and the targeting sequence or region of complementarity is 24 nucleotides in length.

In some embodiments, an oligonucleotide herein comprises a targeting sequence or a region of complementarity (e.g., an antisense strand or a guide strand of a double-stranded oligonucleotide) that is fully complementary to a KHK target sequence. In some embodiments, the targeting sequence or region of complementarity is partially complementary to a KHK target sequence. In some embodiments, the targeting sequence or region of complementarity has up to 3 nucleotide mismatches to a KHK target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to a sequence of KHK. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to a sequence of KHK. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to a sequence of any one of SEQ ID NOs: 4-387. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to a sequence of KHK. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to nucleotides 1-19 of a sequence of any one of SEQ ID NOs: 4-387. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to the sequence set forth in SEQ ID NO: 39. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 39. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to the sequence set forth in SEQ ID NO: 102. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 102. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to the sequence set forth in SEQ ID NO: 104. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 104. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to the sequence set forth in SEQ ID NO: 107. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 107. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to the sequence set forth in SEQ ID NO: 191. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 191. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to the sequence set forth in SEQ ID NO: 269. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 269. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to a sequence of any one of SEQ ID NOs: 4-387. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to nucleotides 1-19 of a sequence of any one of SEQ ID NOs: 4-387. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to the sequence set forth in SEQ ID NO: 39. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 39. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to the sequence set forth in SEQ ID NO: 102. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 102. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to the sequence set forth in SEQ ID NO: 104. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 104. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to the sequence set forth in SEQ ID NO: 107. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 107. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to the sequence set forth in SEQ ID NO: 191. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 191. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to the sequence set forth in SEQ ID NO: 269. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to nucleotides 1-19 of the sequence set forth in SEQ ID NO: 269. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to a sequence of any one of SEQ ID NOs: 872-878 and 886-911. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to a sequence of any one of SEQ ID NOs: 872-878 and 886-911. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is fully complementary to a sequence of any one of SEQ ID NOs: 887, 891, 892, 894, 897 and 909. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is partially complementary to a sequence of any one of SEQ ID NOs: 887, 891, 892, 894, 897 and 909.

In some embodiments, the oligonucleotide herein comprises a targeting sequence or region of complementarity that is complementary to a contiguous sequence of nucleotides comprising a KHK mRNA, wherein the contiguous sequence of nucleotides is about 12 to about 30 nucleotides in length (e.g., 12 to 30, 12 to 28, 12 to 26, 12 to 24, 12 to 20, 12 to 18, 12 to 16, 14 to 22, 16 to 20, 18 to 20 or 18 to 19 nucleotides in length). In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is complementary to a contiguous sequence of nucleotides comprising a KHK mRNA, wherein the contiguous sequence of nucleotides is 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 nucleotides in length. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is complementary to a contiguous sequence of nucleotides comprising a KHK mRNA, wherein the contiguous sequence of nucleotides is 19 nucleotides in length. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity that is complementary to a contiguous sequence of nucleotides comprising a KHK mRNA, wherein the contiguous sequence of nucleotides is 20 nucleotides in length. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 4-387, optionally wherein the contiguous sequence of nucleotides is 19 nucleotides in length. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 4-387, wherein the contiguous sequence of nucleotides is 19 nucleotides in length. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 4-387, wherein the contiguous sequence of nucleotides is 20 nucleotides in length. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 872-878 and 886-911, optionally wherein the contiguous sequence of nucleotides is 19 nucleotides in length. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 872-878 and 886-911, wherein the contiguous sequence of nucleotides is 19 nucleotides in length. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 872-878 and 886-911, wherein the contiguous sequence of nucleotides is 20 nucleotides in length. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909, optionally wherein the contiguous sequence of nucleotides is 19 nucleotides in length. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909, wherein the contiguous sequence of nucleotides is 19 nucleotides in length. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909, wherein the contiguous sequence of nucleotides is 20 nucleotides in length.

In some embodiments, a targeting sequence or region of complementarity of an oligonucleotide is provided that is complementary to contiguous nucleotides of a sequence as set forth in any one of SEQ ID NOs: 4-387 and spans the entire length of an antisense strand. In some embodiments, a targeting sequence or region of complementarity of an oligonucleotide is provided that is complementary to contiguous nucleotides of nucleotides 1-19 a sequence as set forth in any one of SEQ ID NOs: 4-387 and spans the entire length of an antisense strand. In some embodiments, a region of complementarity of an oligonucleotide is provided that is complementary to contiguous nucleotides of a sequence as set forth in any one of SEQ ID NOs: 4-387 and spans a portion of the entire length of an antisense strand. In some embodiments, a region of complementarity of an oligonucleotide is provided that is complementary to contiguous nucleotides of nucleotides 1-19 a sequence as set forth in any one of SEQ ID NOs: 4-387 and spans a portion of the entire length of an antisense strand. In some embodiments, an oligonucleotide herein comprises a region of complementarity (e.g., on an antisense strand of a dsRNA) that is at least partially (e.g., fully) complementary to a contiguous stretch of nucleotides spanning nucleotides 1-19 of a sequence as set forth in any one of SEQ ID NOs: 4-387. In some embodiments, an oligonucleotide herein comprises a region of complementarity (e.g., on an antisense strand of a dsRNA) that is at least partially (e.g., fully) complementary to a contiguous stretch of nucleotides spanning nucleotides 1-20 of a sequence as set forth in any one of SEQ ID NOs: 4-387. In some embodiments, a targeting sequence or region of complementarity of an oligonucleotide is provided that is complementary to contiguous nucleotides of a sequence as set forth in any one of SEQ ID NOs: 872-878 and 886-911 and spans the entire length of an antisense strand. In some embodiments, a region of complementarity of an oligonucleotide that is complementary to contiguous nucleotides of a sequence as set forth in any one of SEQ ID NOs: 872-878 and 886-911 and spans a portion of the entire length of an antisense strand. In some embodiments, an oligonucleotide herein comprises a region of complementarity (e.g., on an antisense strand of a dsRNA) that is at least partially (e.g., fully) complementary to a contiguous stretch of nucleotides spanning nucleotides 1-19 of a sequence as set forth in any one of SEQ ID NOs: 872-878 and 886-911. In some embodiments, an oligonucleotide herein comprises a region of complementarity (e.g., on an antisense strand of a dsRNA) that is at least partially (e.g., fully) complementary to a contiguous stretch of nucleotides spanning nucleotides 1-20 of a sequence as set forth in any one of SEQ ID NOs: 872-878 and 886-911. In some embodiments, a targeting sequence or region of complementarity of an oligonucleotide is provided that is complementary to contiguous nucleotides of a sequence as set forth in any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909 and spans the entire length of an antisense strand. In some embodiments, a region of complementarity of an oligonucleotide is provided that is complementary to contiguous nucleotides of a sequence as set forth in any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909 and spans a portion of the entire length of an antisense strand. In some embodiments, an oligonucleotide herein comprises a region of complementarity (e.g., on an antisense strand of a dsRNA) that is at least partially (e.g., fully) complementary to a contiguous stretch of nucleotides spanning nucleotides 1-19 of a sequence as set forth in any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909. In some embodiments, an oligonucleotide herein comprises a region of complementarity (e.g., on an antisense strand of a dsRNA) that is at least partially (e.g., fully) complementary to a contiguous stretch of nucleotides spanning nucleotides 1-20 of a sequence as set forth in any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909.

In some embodiments, an oligonucleotide herein comprises a targeting sequence or region of complementarity having one or more base pair (bp) mismatches with the corresponding KHK target sequence. In some embodiments, the targeting sequence or region of complementarity may have up to about 1, up to about 2, up to about 3, up to about 4, up to about 5, etc. mismatches with the corresponding KHK target sequence provided that the ability of the targeting sequence or region of complementarity to bind or anneal to the KHK mRNA under appropriate hybridization conditions and/or the ability of the oligonucleotide to inhibit KHK expression is maintained. Alternatively, the targeting sequence or region of complementarity may have no more than 1, no more than 2, no more than 3, no more than 4, or no more than 5 mismatches with the corresponding KHK target sequence provided that the ability of the targeting sequence or region of complementarity to bind or anneal to the KHK mRNA under appropriate hybridization conditions and/or the ability of the oligonucleotide to inhibit KHK expression is maintained. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity having 1 mismatch with the corresponding target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity having 2 mismatches with the corresponding target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity having 3 mismatches with the corresponding target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity having 4 mismatches with the corresponding target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity having 5 mismatches with the corresponding target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity having more than one mismatch (e.g., 2, 3, 4, 5 or more mismatches) with the corresponding target sequence, wherein at least 2 (e.g., all) of the mismatches are positioned consecutively (e.g., 2, 3, 4, 5 or more mismatches in a row), or wherein the mismatches are interspersed throughout the targeting sequence or region of complementarity. In some embodiments, the oligonucleotide comprises a targeting sequence or region of complementarity having more than one mismatch (e.g., 2, 3, 4, 5 or more mismatches) with the corresponding target sequence, wherein at least 2 (e.g., all) of the mismatches are positioned consecutively (e.g., 2, 3, 4, 5 or more mismatches in a row), or wherein at least one or more non-mismatched base pair is located between the mismatches, or a combination thereof. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 4-387, wherein the targeting sequence or region of complementarity may have up to about 1, up to about 2, up to about 3, up to about 4, up to about 5, etc. mismatches with the corresponding KHK target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of nucleotides 1-19 of any one of SEQ ID NOs: 4-387, wherein the targeting sequence or region of complementarity may have up to about 1, up to about 2, up to about 3, up to about 4, up to about 5, etc. mismatches with the corresponding KHK target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 4-387, wherein the targeting sequence or region of complementarity may have no more than 1, no more than 2, no more than 3, no more than 4, or no more than 5 mismatches with the corresponding KHK target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of nucleotides 1-19 of any one of SEQ ID NOs: 4-387, wherein the targeting sequence or region of complementarity may have no more than 1, no more than 2, no more than 3, no more than 4, or no more than 5 mismatches with the corresponding KHK target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 872-878 and 886-911, wherein the targeting sequence or region of complementarity may have up to about 1, up to about 2, up to about 3, up to about 4, up to about 5, etc. mismatches with the corresponding KHK target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 872-878 and 886-911, wherein the targeting sequence or region of complementarity may have no more than 1, no more than 2, no more than 3, no more than 4, or no more than 5 mismatches with the corresponding KHK target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909, wherein the targeting sequence or region of complementarity may have up to about 1, up to about 2, up to about 3, up to about 4, up to about 5, etc. mismatches with the corresponding KHK target sequence. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909, wherein the targeting sequence or region of complementarity may have no more than 1, no more than 2, no more than 3, no more than 4, or no more than 5 mismatches with the corresponding KHK target sequence.

Types of Oligonucleotides

A variety of oligonucleotide types and/or structures are useful for targeting KHK in the methods herein including, but not limited to, RNAi oligonucleotides, antisense oligonucleotides, miRNAs, etc. Any of the oligonucleotide types described herein or elsewhere are contemplated for use as a framework to incorporate a KHK targeting sequence herein for the purposes of inhibiting KHK expression.

In some embodiments, the oligonucleotides herein inhibit KHK expression by engaging with RNA interference (RNAi) pathways upstream or downstream of Dicer involvement. For example, RNAi oligonucleotides have been developed with each strand having sizes of about 19-25 nucleotides with at least one 3′ overhang of 1 to 5 nucleotides (see, e.g., U.S. Pat. No. 8,372,968). Longer oligonucleotides also have been developed that are processed by Dicer to generate active RNAi products (see, e.g., U.S. Pat. No. 8,883,996). Further work produced extended dsRNAs where at least one end of at least one strand is extended beyond a duplex targeting region, including structures where one of the strands includes a thermodynamically stabilizing tetraloop structure (see, e.g., U.S. Pat. Nos. 8,513,207 and 8,927,705, as well as Intl. Patent Application Publication No. WO 2010/033225). Such structures may include single-stranded (ss) extensions (on one or both sides of the molecule) as well as double-stranded (ds) extensions.

In some embodiments, the oligonucleotides herein engage with the RNAi pathway downstream of the involvement of Dicer (e.g., Dicer cleavage). In some embodiments, the oligonucleotides described herein are Dicer substrates. In some embodiments, upon endogenous Dicer processing, double-stranded nucleic acids of 19-23 nucleotides in length capable of reducing KHK expression are produced. In some embodiments, the oligonucleotide has an overhang (e.g., of 1, 2, or 3 nucleotides in length) in the 3′ end of the sense strand. In some embodiments, the oligonucleotide has an overhang (e.g., of 1, 2, or 3 nucleotides in length) in the 3′ end of the antisense strand. In some embodiments, the oligonucleotide (e.g., siRNA) comprises a 21-nucleotide guide strand that is antisense to a target RNA and a complementary passenger strand, in which both strands anneal to form a 19-bp duplex and 2 nucleotide overhangs at either or both 3′ ends. Longer oligonucleotide designs also are available including oligonucleotides having a guide strand of 23 nucleotides and a passenger strand of 21 nucleotides, where there is a blunt end on the right side of the molecule (3′ end of passenger strand/5′ end of guide strand) and a two nucleotide 3′-guide strand overhang on the left side of the molecule (5′ end of the passenger strand/3′ end of the guide strand). In such molecules, there is a 21 bp duplex region. See, e.g., U.S. Pat. Nos. 9,012,138; 9,012,621 and 9,193,753.

In some embodiments, the oligonucleotides herein comprise sense and antisense strands that are both in the range of about 17 to 36 (e.g., 17 to 36, 20 to 25 or 21-23) nucleotides in length. In some embodiments, the oligonucleotides described herein comprise an antisense strand of 19-30 nucleotides in length and a sense strand of 19-50 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand. In some embodiments, an oligonucleotide herein comprises a sense and antisense strand that are both in the range of about 19-22 nucleotides in length. In some embodiments, the sense and antisense strands are of equal length. In some embodiments, an oligonucleotide comprises sense and antisense strands, such that there is a 3′-overhang on either the sense strand or the antisense strand, or both the sense and antisense strand. In some embodiments, for oligonucleotides that have sense and antisense strands that are both in the range of about 21-23 nucleotides in length, a 3′ overhang on the sense, antisense, or both sense and antisense strands is 1 or 2 nucleotides in length. In some embodiments, the oligonucleotide has a guide strand of 22 nucleotides and a passenger strand of 20 nucleotides, where there is a blunt end on the right side of the molecule (3′ end of passenger strand/5′ end of guide strand) and a 2 nucleotide 3′-guide strand overhang on the left side of the molecule (5′ end of the passenger strand/3′ end of the guide strand). In such molecules, there is a 20 bp duplex region.

Other oligonucleotide designs for use with the compositions and methods herein include: 16-mer siRNAs (see, e.g., NUCLEIC ACIDS IN CHEMISTRY AND BIOLOGY. Blackburn (ed.), Royal Society of Chemistry, 2006), shRNAs (e.g., having 19 bp or shorter stems; see, e.g., Moore et al. (2010) METHODS MOL. BIOL. 629:139-156), blunt siRNAs (e.g., of 19 bps in length; see, e.g., Kraynack & Baker (2006) RNA 12:163-176), asymmetrical siRNAs (aiRNA; see, e.g., Sun et al. (2008) NAT. BIOTECHNOL. 26:1379-1382), asymmetric shorter-duplex siRNA (see, e.g., Chang et al. (2009) MOL. THER. 17:725-32), fork siRNAs (see, e.g., Hohjoh (2004) FEBS LETT. 557:193-198), ss siRNAs (Elsner (2012) NAT. BIOTECHNOL. 30:1063), dumbbell-shaped circular siRNAs (see, e.g., Abe et al. (2007) J. AM. CHEM. SOC. 129:15108-09), and small internally segmented interfering RNA (siRNA; see, e.g., Bramsen et al. (2007) NUCLEIC ACIDS RES. 35:5886-97). Further non-limiting examples of an oligonucleotide structures that may be used in some embodiments to reduce or inhibit the expression of KHK are microRNA (miRNA), short hairpin RNA (shRNA) and short siRNA (see, e.g., Hamilton et al. (2002) EMBO J. 21:4671-79; see also, US Patent Application Publication No. 2009/0099115).

Still, in some embodiments, an oligonucleotide for reducing or inhibiting KHK expression herein is single-stranded (ss). Such structures may include but are not limited to single-stranded RNAi molecules. Recent efforts have demonstrated the activity of ss RNAi molecules (see, e.g., Matsui et al. (2016) MOL. THER. 24:946-955). However, in some embodiments, oligonucleotides herein are antisense oligonucleotides (ASOs). An antisense oligonucleotide is a single-stranded oligonucleotide that has a nucleobase sequence which, when written in the 5′ to 3′ direction, comprises the reverse complement of a targeted segment of a particular nucleic acid and is suitably modified (e.g., as a gapmer) so as to induce RNaseH-mediated cleavage of its target RNA in cells or (e.g., as a mixmer) so as to inhibit translation of the target mRNA in cells. ASOs for use herein may be modified in any suitable manner known in the art including, for example, as shown in U.S. Pat. No. 9,567,587 (including, e.g., length, sugar moieties of the nucleobase (pyrimidine, purine), and alterations of the heterocyclic portion of the nucleobase). Further, ASOs have been used for decades to reduce expression of specific target genes (see, e.g., Bennett et al. (2017) ANNU. REV. PHARMACOL. 57:81-105).

In some embodiments, the antisense oligonucleotide shares a region of complementarity with KHK mRNA. In some embodiments, the antisense oligonucleotide targets SEQ ID NO: 1. In some embodiments, the antisense oligonucleotide targets SEQ ID NO: 2. In some embodiments, the antisense oligonucleotide targets SEQ ID NO: 3. In some embodiments, the antisense oligonucleotide is 15-50 nucleotides in length. In some embodiments, the antisense oligonucleotide is 15-25 nucleotides in length. In some embodiments, the antisense oligonucleotide is 22 nucleotides in length. In some embodiments, the antisense oligonucleotide is complementary to any one of SEQ ID NOs: 4-387. In some embodiments, the antisense oligonucleotide is complementary to nucleotides 1-19 of any one of SEQ ID NOs: 4-387. In some embodiments, the antisense oligonucleotide is at least 15 contiguous nucleotides in length. In some embodiments, the antisense oligonucleotide is at least 19 contiguous nucleotides in length. In some embodiments, the antisense oligonucleotide is at least 20 contiguous nucleotides in length. In some embodiments, the antisense oligonucleotide differs by 1, 2, or 3 nucleotides from the target sequence.

Double-Stranded Oligonucleotides

In some aspects, the disclosure provides double-stranded (ds) RNAi oligonucleotides for targeting KHK mRNA and inhibiting KHK expression (e.g., via the RNAi pathway) comprising a sense strand (also referred to herein as a passenger strand) and an antisense strand (also referred to herein as a guide strand). In some embodiments, the sense strand and antisense strand are separate strands and are not covalently linked. In some embodiments, the sense strand and antisense strand are covalently linked. In some embodiments, the sense strand and antisense strand form a duplex region, wherein the sense strand and antisense strand, or a portion thereof, binds with one another in a complementary fashion (e.g., by Watson-Crick base pairing).

In some embodiments, the sense strand has a first region (R1) and a second region (R2), wherein R2 comprises a first subregion (S1), a tetraloop (L) or triloop (triL), and a second subregion (S2), wherein L or triL is located between S1 and S2, and wherein S1 and S2 form a second duplex (D2). D2 may have various length. In some embodiments, D2 is about 1-6 bp in length. In some embodiments, D2 is 2-6, 3-6, 4-6, 5-6, 1-5, 2-5, 3-5 or 4-5 bp in length. In some embodiments, D2 is 1, 2, 3, 4, 5 or 6 bp in length. In some embodiments, D2 is 6 bp in length.

In some embodiments, R1 of the sense strand and the antisense strand form a first duplex (D1). In some embodiments, D1 is at least about 15 (e.g., at least 15, at least 16, at least 17, at least 18, at least 19, at least 20 or at least 21) nucleotides in length. In some embodiments, D1 is in the range of about 12 to 30 nucleotides in length (e.g., 12 to 30, 12 to 27, 15 to 22, 18 to 22, 18 to 25, 18 to 27, 18 to 30 or 21 to 30 nucleotides in length). In some embodiments, D1 is at least 12 nucleotides in length (e.g., at least 12, at least 15, at least 20, at least 25, or at least 30 nucleotides in length). In some embodiments, D1 is 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 nucleotides in length. In some embodiments, D1 is 20 nucleotides in length. In some embodiments, D1 comprising sense strand and antisense strand does not span the entire length of the sense strand and/or antisense strand. In some embodiments, D1 comprising the sense strand and antisense strand spans the entire length of either the sense strand or antisense strand or both. In certain embodiments, D1 comprising the sense strand and antisense strand spans the entire length of both the sense strand and the antisense strand.

In some embodiments, a dsRNAi provided herein comprises a sense strand having a sequence of any one of SEQ ID NOs: 4-387; and an antisense strand comprising a complementary sequence selected from SEQ ID NOs: 388-771 as is arranged Table 2.

In some embodiments, a dsRNAi oligonucleotide comprises a sense strand and an antisense strand comprising nucleotide sequences selected from:

• • (a) SEQ ID NOs: 886 and 912, respectively;
  • (b) SEQ ID NOs: 887 and 913, respectively;
  • (c) SEQ ID NOs: 910 and 937, respectively;
  • (d) SEQ ID NOs: 888 and 914, respectively;
  • (e) SEQ ID NOs: 889 and 915, respectively;
  • (f) SEQ ID NOs: 890 and 916, respectively;
  • (g) SEQ ID NOs: 891 and 917, respectively;
  • (h) SEQ ID NOs: 877 and 884, respectively;
  • (i) SEQ ID NOs: 878 and 930, respectively;
  • (j) SEQ ID NOs: 876 and 883, respectively;
  • (k) SEQ ID NOs: 875 and 882, respectively;
  • (l) SEQ ID NOs: 892 and 918, respectively;
  • (m) SEQ ID NOs: 893 and 919, respectively;
  • (n) SEQ ID NOs: 894 and 920, respectively;
  • (o) SEQ ID NOs: 904 and 931, respectively;
  • (p) SEQ ID NOs: 895 and 921, respectively;
  • (q) SEQ ID NOs: 905 and 932, respectively;
  • (r) SEQ ID NOs: 896 and 922, respectively;
  • (s) SEQ ID NOs: 911 and 938, respectively;
  • (t) SEQ ID NOs: 906 and 933, respectively;
  • (u) SEQ ID NOs: 897 and 923, respectively;
  • (v) SEQ ID NOs: 907 and 934, respectively;
  • (w) SEQ ID NOs: 908 and 935, respectively;
  • (x) SEQ ID NOs: 903 and 929, respectively;
  • (y) SEQ ID NOs: 901 and 927, respectively;
  • (z) SEQ ID NOs: 874 and 881, respectively;
  • (aa) SEQ ID NOs: 902 and 928, respectively;
  • (bb) SEQ ID NOs: 873 and 880, respectively;
  • (cc) SEQ ID NOs: 872 and 879, respectively;
  • (dd) SEQ ID NOs: 898 and 924, respectively;
  • (ee) SEQ ID NOs: 899 and 925, respectively (ff) SEQ ID NOs: 900 and 926, respectively; and
  • (gg) SEQ ID NOs: 909 and 936, respectively.

In some embodiments, a dsRNAi oligonucleotide comprises a sense strand and an antisense strand comprising nucleotide sequences selected from:

• • (a) SEQ ID NOs: 887 and 913, respectively;
  • (b) SEQ ID NOs: 891 and 917, respectively;
  • (c) SEQ ID NOs: 892 and 918, respectively;
  • (d) SEQ ID NOs: 894 and 920, respectively;
  • (e) SEQ ID NOs: 897 and 923, respectively; and
  • (f) SEQ ID NOs: 909 and 936, respectively.

In some embodiments, the sense strand comprises the sequence of SEQ ID NO: 887 and the antisense strand comprises the sequence of SEQ ID NO: 913. In some embodiments, the sense strand comprises the sequence of SEQ ID NO: 891 and the antisense strand comprises the sequence of SEQ ID NO: 917. In some embodiments, the sense strand comprises the sequence of SEQ ID NO: 892 and the antisense strand comprises the sequence of SEQ ID NO: 918. In some embodiments, the sense strand comprises the sequence of SEQ ID NO: 894 and the antisense strand comprises the sequence of SEQ ID NO: 920. In some embodiments, the sense strand comprises the sequence of SEQ ID NO: 897 and the antisense strand comprises the sequence of SEQ ID NO: 923. In some embodiments, the sense strand comprises the sequence of SEQ ID NO: 909 and the antisense strand comprises the sequence of SEQ ID NO: 936.

It should be appreciated that, in some embodiments, sequences presented in the Sequence Listing may be referred to in describing the structure of an oligonucleotide (e.g., a dsRNAi oligonucleotide) or other nucleic acid. In such embodiments, the actual oligonucleotide or other nucleic acid may have one or more alternative nucleotides (e.g., an RNA counterpart of a DNA nucleotide or a DNA counterpart of an RNA nucleotide) and/or one or more modified nucleotides and/or one or more modified internucleotide linkages and/or one or more other modification when compared with the specified sequence while retaining essentially same or similar complementary properties as the specified sequence.

In some embodiments, a dsRNAi oligonucleotide herein comprises a 25-nucleotide sense strand and a 27-nucleotide antisense strand that when acted upon by a Dicer enzyme results in an antisense strand that is incorporated into the mature RISC. In some embodiments, the sense strand of the dsRNA is longer than 27 nucleotides (e.g., 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50 nucleotides). In some embodiments, the sense strand of the dsRNA is longer than 25 nucleotides (e.g., 26, 27, 28, 29 or 30 nucleotides). In some embodiments, the sense strand of the dsRNA comprises a nucleotide sequence selected from SEQ ID NOs: 4-387, wherein the nucleotide sequence is longer than 27 nucleotides (e.g., 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 or 50 nucleotides). In some embodiments, the sense strand of the dsRNA comprises a nucleotide sequence selected from SEQ ID NOs: 4-387, wherein the nucleotide sequence is longer than 25 nucleotides (e.g., 26, 27, 28, 29 or 30 nucleotides).

In some embodiments, oligonucleotides herein have one 5′ end that is thermodynamically less stable when compared to the other 5′ end. In some embodiments, an asymmetric oligonucleotide is provided that includes a blunt end at the 3′ end of a sense strand and a 3′-overhang at the 3′ end of an antisense strand. In some embodiments, the 3′-overhang on the antisense strand is about 1-8 nucleotides in length (e.g., 1, 2, 3, 4, 5, 6, 7 or 8 nucleotides in length). Typically, a dsRNAi oligonucleotide has a two-nucleotide overhang on the 3′ end of the antisense (guide) strand. However, other overhangs are possible. In some embodiments, an overhang is a 3′-overhang comprising a length of between 1 and 6 nucleotides, optionally 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 6, 3 to 5, 3 to 4, 4 to 6, 4 to 5, 5 to 6 nucleotides, or 1, 2, 3, 4, 5 or 6 nucleotides. However, in some embodiments, the overhang is a 5′-overhang comprising a length of between 1 and 6 nucleotides, optionally 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 6, 3 to 5, 3 to 4, 4 to 6, 4 to 5, 5 to 6 nucleotides, or 1, 2, 3, 4, 5 or 6 nucleotides. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 4-387, and a 5′-overhang comprising a length of between 1 and 6 nucleotides. In some embodiments, the oligonucleotide comprises a sense strand comprising a nucleotide sequence selected from SEQ ID NOs: 4-387, wherein the oligonucleotide comprises a 5′-overhang comprising a length of between 1 and 6 nucleotides. In some embodiments, the oligonucleotide comprises an antisense strand comprising a nucleotide sequence selected from SEQ ID NOs: 388-771, wherein the oligonucleotide comprises a 5′-overhang comprising a length of between 1 and 6 nucleotides. In some embodiments, the oligonucleotide comprises a sense strand comprising a nucleotide sequence selected from SEQ ID NOs: 4-387 and antisense strand comprising a nucleotide sequence selected from SEQ ID NOs: 388-771, wherein the oligonucleotide comprises a 5′-overhang comprising a length of between 1 and 6 nucleotides.

In some embodiments, two terminal nucleotides on the 3′ end of an antisense strand are modified. In some embodiments, the two terminal nucleotides on the 3′ end of the antisense strand are complementary with the target mRNA (e.g., KHK mRNA). In some embodiments, the two terminal nucleotides on the 3′ end of the antisense strand are not complementary with the target mRNA. In some embodiments, the two terminal nucleotides on the 3′ end of the antisense strand of a dsRNAi oligonucleotide herein are unpaired. In some embodiments, the two terminal nucleotides on the 3′ end of the antisense strand of a dsRNAi oligonucleotide herein comprise an unpaired GG. In some embodiments, the two terminal nucleotides on the 3′ end of an antisense strand of a dsRNAi oligonucleotide herein are not complementary to the target mRNA. In some embodiments, two terminal nucleotides on each 3′ end of a dsRNAi oligonucleotide are GG. Typically, one or both of the two terminal GG nucleotides on each 3′ end of a double-stranded oligonucleotide is not complementary with the target mRNA. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 4-387, wherein the two terminal nucleotides on the 3′ end of the antisense strand of a dsRNAi oligonucleotide herein comprise an unpaired GG. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of nucleotides 1-19 of any one of SEQ ID NOs: 4-387, wherein the two terminal nucleotides on the 3′ end of the antisense strand of a dsRNAi oligonucleotide herein comprise an unpaired GG. In some embodiments, the oligonucleotide comprises an antisense strand comprising a nucleotide sequence selected from SEQ ID NOs: 388-771, wherein the two terminal nucleotides on the 3′ end of the antisense strand of a dsRNAi oligonucleotide herein comprise an unpaired GG. In some embodiments, the oligonucleotide comprises a sense strand comprising a nucleotide sequence selected from SEQ ID NOs: 4-387 and antisense strand comprising a nucleotide sequence selected from SEQ ID NOs: 388-771, wherein the two terminal nucleotides on the 3′ end of the antisense strand of a dsRNAi oligonucleotide herein comprise an unpaired GG.

In some embodiments, there is one or more (e.g., 1, 2, 3, 4 or 5) mismatch(es) between a sense and antisense strand. If there is more than one mismatch between a sense and antisense strand, they may be positioned consecutively (e.g., 2, 3 or more in a row), or interspersed throughout the region of complementarity. In some embodiments, the 3′ end of the sense strand contains one or more mismatches. In some embodiments, two mismatches are incorporated at the 3′ end of the sense strand. In some embodiments, base mismatches, or destabilization of segments at the 3′ end of the sense strand of the dsRNAi oligonucleotide improves or increases the potency of the dsRNAi oligonucleotide. In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein there is one or more (e.g., 1, 2, 3, 4 or 5) mismatch(es) between the sense and antisense strands.

Antisense Strands

In some embodiments, an antisense strand of a dsRNAi oligonucleotide is referred to as a “guide strand.” For example, an antisense strand that engages with RNA-induced silencing complex (RISC) and binds to an Argonaute protein such as Ago2, or engages with or binds to one or more similar factors, and directs silencing of a target gene, as the antisense strand is referred to as a guide strand. In some embodiments, a sense strand complementary to a guide strand may be referred to as a “passenger strand.”

In some embodiments, a dsRNAi oligonucleotide herein comprises an antisense strand of up to about 50 nucleotides in length (e.g., up to 50, up to 40, up to 35, up to 30, up to 27, up to 25, up to 21, up to 19, up to 17 or up to 12 nucleotides in length). In some embodiments, a dsRNAi oligonucleotide comprises an antisense strand of at least about 12 nucleotides in length (e.g., at least 12, at least 15, at least 19, at least 21, at least 22, at least 25, at least 27, at least 30, at least 35 or at least 38 nucleotides in length). In some embodiments, a dsRNAi oligonucleotide comprises an antisense strand in a range of about 12 to about 40 (e.g., 12 to 40, 12 to 36, 12 to 32, 12 to 28, 15 to 40, 15 to 36, 15 to 32, 15 to 28, 17 to 22, 17 to 25, 19 to 27, 19 to 30, 20 to 40, 22 to 40, 25 to 40 or 32 to 40) nucleotides in length. In some embodiments, a dsRNAi oligonucleotide comprises antisense strand of 15 to 30 nucleotides in length. In some embodiments, an antisense strand of any one of the dsRNAi oligonucleotides disclosed herein is of 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40 nucleotides in length. In some embodiments, a dsRNAi oligonucleotide comprises an antisense strand of 22 nucleotides in length.

In some embodiments, a dsRNAi oligonucleotide disclosed herein for targeting KHK comprises an antisense strand comprising or consisting of a sequence as set forth in any one of SEQ ID NOs: 388-771. In some embodiments, a dsRNAi oligonucleotide herein comprises an antisense strand comprising at least about 12 (e.g., at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22 or at least 23) contiguous nucleotides of a sequence as set forth in any one of SEQ ID NOs: 388-771. In some embodiments, a dsRNAi oligonucleotide disclosed herein for targeting KHK comprises an antisense strand comprising or consisting of a sequence as set forth in any one of SEQ ID NOs: 879-885 and 912-938. In some embodiments, a dsRNAi oligonucleotide herein comprises an antisense strand comprising at least about 12 (e.g., at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22 or at least 23) contiguous nucleotides of a sequence as set forth in any one of SEQ ID NOs: 879-885 and 912-938. In some embodiments, a dsRNAi oligonucleotide disclosed herein for targeting KHK comprises an antisense strand comprising or consisting of a sequence as set forth in any one of SEQ ID NOs: 913, 917, 918, 920, 923 and 936. In some embodiments, a dsRNAi oligonucleotide herein comprises an antisense strand comprising at least about 12 (e.g., at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22 or at least 23) contiguous nucleotides of a sequence as set forth in any one of SEQ ID NOs: 913, 917, 918, 920, 923 and 936.

In some embodiments, a dsRNAi oligonucleotide herein comprises an antisense strand comprising a nucleotide sequence selected from SEQ ID NOs: 948-953.

Sense Strands

In some embodiments, a dsRNAi oligonucleotide disclosed herein for targeting KHK mRNA and inhibiting KHK expression comprises a sense strand sequence as set forth in any one of SEQ ID NOs: 4-387. In some embodiments, a dsRNAi oligonucleotide has a sense strand that comprises at least about 12 (e.g., at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22 or at least 23) contiguous nucleotides of a sequence as set forth in any one of SEQ ID NOs: 4-387. In some embodiments, a dsRNAi oligonucleotide disclosed herein for targeting KHK mRNA and inhibiting KHK expression comprises a sense strand sequence as set forth in any one of SEQ ID NOs: 872-878 and 886-911. In some embodiments, a dsRNAi oligonucleotide has a sense strand that comprises at least about 12 (e.g., at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22 or at least 23) contiguous nucleotides of a sequence as set forth in in any one of SEQ ID NOs: 872-878 and 886-911. In some embodiments, a dsRNAi oligonucleotide disclosed herein for targeting KHK mRNA and inhibiting KHK expression comprises a sense strand sequence as set forth in any one of SEQ ID NOs: 887, 891, 892, 894, 897 and 909. In some embodiments, a dsRNAi oligonucleotide has a sense strand that comprises at least about 12 (e.g., at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22 or at least 23) contiguous nucleotides of a sequence as set forth in any one of SEQ ID NOs: 887, 891, 892, 894, 897 and 909.

In some embodiments, a dsRNAi oligonucleotide herein comprises a sense strand (or passenger strand) of up to about 50 nucleotides in length (e.g., up to 50, up to 40, up to 36, up to 30, up to 27, up to 25, up to 21, up to 19, up to 17 or up to 12 nucleotides in length). In some embodiments, a dsRNAi oligonucleotide may have a sense strand of at least about 12 nucleotides in length (e.g., at least 12, at least 15, at least 19, at least 21, at least 25, at least 27, at least 30, at least 36 or at least 38 nucleotides in length). In some embodiments, an oligonucleotide may have a sense strand in a range of about 12 to about 50 (e.g., 12 to 50, 12 to 40, 12 to 36, 12 to 32, 12 to 28, 15 to 40, 15 to 36, 15 to 32, 15 to 28, 17 to 21, 17 to 25, 19 to 27, 19 to 30, 20 to 40, 22 to 40, 25 to 40 or 32 to 40) nucleotides in length. In some embodiments, a dsRNAi oligonucleotide comprises a sense strand of 15 to 50 nucleotides in length. In some embodiments, a dsRNAi oligonucleotide comprises a sense strand of 18 to 36 nucleotides in length. In some embodiments, an oligonucleotide may have a sense strand of 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides in length. In some embodiments, a dsRNAi oligonucleotide comprises a sense strand of 36 nucleotides in length.

In some embodiments, a sense strand comprises a stem-loop structure at its 3′ end. In some embodiments, the stem-loop is formed by intrastrand base pairing. In some embodiments, a sense strand comprises a stem-loop structure at its 5′ end. In some embodiments, a stem is a duplex of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 nucleotides in length. In some embodiments, a stem-loop provides the dsRNAi oligonucleotide protection against degradation (e.g., enzymatic degradation), facilitates or improves targeting and/or delivery to a target cell, tissue, or organ (e.g., the liver), or both. For example, in some embodiments, the loop of a stem-loop provides nucleotides comprising one or more modifications that facilitate, improve, or increase targeting to a target mRNA (e.g., a KHK mRNA), inhibition of target gene expression (e.g., KHK expression), and/or delivery to a target cell, tissue, or organ (e.g., the liver), or a combination thereof. In some embodiments, the stem-loop itself or modification(s) to the stem-loop do not substantially affect the inherent gene expression inhibition activity of the dsRNAi oligonucleotide, but facilitates, improves, or increases stability (e.g., provides protection against degradation) and/or delivery of the oligonucleotide to a target cell, tissue, or organ (e.g., the liver). In certain embodiments, a dsRNAi oligonucleotide comprises a sense strand comprising (e.g., at its 3′ end) a stem-loop set forth as: S1-L-S2, in which S1 is complementary to S2, and in which L forms a single-stranded loop between S1 and S2 of up to about 10 nucleotides in length (e.g., 3, 4, 5, 6, 7, 8, 9 or 10 nucleotides in length). In some embodiments, the loop (L) is 3 nucleotides in length. In some embodiments, the loop (L) is 4 nucleotides in length. In some embodiments, an oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 4-387, and the oligonucleotide comprises a sense strand comprising (e.g., at its 3′ end) a stem-loop set forth as: S1-L-S2, in which S1 is complementary to S2, and in which L forms a single-stranded loop between S1 and S2 of up to about 10 nucleotides in length (e.g., 3, 4, 5, 6, 7, 8, 9 or 10 nucleotides in length). In some embodiments, an oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides 1-19 of any one of SEQ ID NOs: 4-387, and the oligonucleotide comprises a sense strand comprising (e.g., at its 3′ end) a stem-loop set forth as: S1-L-S2, in which 51 is complementary to S2, and in which L forms a single-stranded loop between S1 and S2 of up to about 10 nucleotides in length (e.g., 3, 4, 5, 6, 7, 8, 9 or 10 nucleotides in length). In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 872-878 and 886-911, and the oligonucleotide comprises a sense strand comprising (e.g., at its 3′ end) a stem-loop set forth as: S1-L-S2, in which 51 is complementary to S2, and in which L forms a single-stranded loop between S1 and S2 of up to about 10 nucleotides in length (e.g., 3, 4, 5, 6, 7, 8, 9 or 10 nucleotides in length). In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909, and the oligonucleotide comprises a sense strand comprising (e.g., at its 3′ end) a stem-loop set forth as: S1-L-S2, in which 51 is complementary to S2, and in which L forms a single-stranded loop between S1 and S2 of up to about 10 nucleotides in length (e.g., 3, 4, 5, 6, 7, 8, 9 or 10 nucleotides in length).

In some embodiments, a loop (L) of a stem-loop having the structure S1-L-S2 as described above is a triloop. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 4-387 and a triloop. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides 1-19 of any one of SEQ ID NOs: 4-387 and a triloop. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 872-878 and 886-911, and a triloop. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909, and a triloop. In some embodiments, the triloop comprises ribonucleotides, deoxyribonucleotides, modified nucleotides, delivery ligands, and combinations thereof.

In some embodiments, a loop (L) of a stem-loop having the structure S1-L-S2 as described above is a tetraloop (e.g., within a nicked tetraloop structure) comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 4-387 and a tetraloop. In some embodiments, a loop (L) of a stem-loop having the structure S1-L-S2 as described above is a tetraloop (e.g., within a nicked tetraloop structure) comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides 1-19 of any one of SEQ ID NOs: 4-387 and a tetraloop. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 872-878 and 886-911, and a tetraloop. In some embodiments, the oligonucleotide comprises a targeting sequence or a region of complementarity that is complementary to a contiguous sequence of nucleotides of any one of SEQ ID NOs: 887, 891, 892, 894, 897, and 909, and a tetraloop. In some embodiments, the tetraloop comprises ribonucleotides, deoxyribonucleotides, modified nucleotides, delivery ligands, and combinations thereof.

In some embodiments, a dsRNAi oligonucleotide herein comprises a sense strand comprising a nucleotide sequence selected from SEQ ID NOs: 942-947.

Duplex Length

In some embodiments, a duplex formed between a sense and antisense strand is at least 12 (e.g., at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, or at least 21) nucleotides in length. In some embodiments, a duplex formed between a sense and antisense strand is in the range of 12-30 nucleotides in length (e.g., 12 to 30, 12 to 27, 12 to 22, 15 to 25, 18 to 30, 18 to 22, 18 to 25, 18 to 27, 18 to 30, 19 to 30 or 21 to 30 nucleotides in length). In some embodiments, a duplex formed between a sense and antisense strand is 12, 13, 14, 15, 16, 17, 18, 19, 29, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 nucleotides in length. In some embodiments, a duplex formed between a sense and antisense strand does not span the entire length of the sense strand and/or antisense strand. In some embodiments, a duplex between a sense and antisense strand spans the entire length of either the sense or antisense strands. In some embodiments, a duplex between a sense and antisense strand spans the entire length of both the sense strand and the antisense strand. In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein a duplex formed between a sense and antisense strand is in the range of 12-30 nucleotides in length (e.g., 12 to 30, 12 to 27, 12 to 22, 15 to 25, 18 to 30, 18 to 22, 18 to 25, 18 to 27, 18 to 30, 19 to 30 or 21 to 30 nucleotides in length).

Oligonucleotide Ends

In some embodiments, a dsRNAi oligonucleotide herein comprises sense and antisense strands, such that there is a 3′-overhang on either the sense strand or the antisense strand, or both the sense and antisense strand. In some embodiments, a dsRNAi oligonucleotide herein comprises sense and antisense strands that are separate strands which form an asymmetric duplex region having an overhang at the 3′ terminus of the antisense strand. In some embodiments, a dsRNAi oligonucleotide provided herein has one 5′end that is thermodynamically less stable compared to the other 5′ end. In some embodiments, an asymmetric dsRNAi oligonucleotide is provided that includes a blunt end at the 3′end of a sense strand and overhang at the 3′ end of the antisense strand. In some embodiments, a 3′ overhang on an antisense strand is 1-8 nucleotides in length (e.g., 1, 2, 3, 4, 5, 6, 7 or 8 nucleotides in length). In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the antisense strand comprises a 3′ overhang of 1-8 nucleotides in length (e.g., 1, 2, 3, 4, 5, 6, 7 or 8 nucleotides in length).

Typically, an oligonucleotide for RNAi has a two (2) nucleotide overhang on the 3′ end of the antisense (guide) strand. However, other overhangs are possible. In some embodiments, an overhang is a 3′ overhang comprising a length of between one and six nucleotides, optionally one to five, one to four, one to three, one to two, two to six, two to five, two to four, two to three, three to six, three to five, three to four, four to six, four to five, five to six nucleotides or one, two, three, four, five or six nucleotides. In some embodiments, the overhang is a 5′ overhang comprising a length of between one and six nucleotides, optionally one to five, one to four, one to three, one to two, two to six, two to five, two to four, two to three, three to six, three to five, three to four, four to six, four to five, five to six nucleotides or one, two, three, four, five or six nucleotides. In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the antisense strand comprises a 5′ overhang of 1-6 nucleotides in length.

In some embodiments, one or more (e.g., 2, 3, 4) terminal nucleotides of the 3′ end or 5′ end of a sense and/or antisense strand are modified. For example, in some embodiments, one or two terminal nucleotides of the 3′ end of the antisense strand are modified. In some embodiments, the last nucleotide at the 3′ end of an antisense strand is modified, e.g., comprises 2′ modification, e.g., a 2′-O-methoxyethyl. In some embodiments, the last one or two terminal nucleotides at the 3′ end of an antisense strand are complementary with the target.

In some embodiments, the last one or two nucleotides at the 3′ end of the antisense strand are not complementary with the target.

In some embodiments, a dsRNAi oligonucleotide herein comprises a stem-loop structure at the 3′ end of the sense strand and comprises two terminal overhang nucleotides at the 3′ end of the antisense strand. In some embodiments, a dsRNAi oligonucleotide herein comprises a nicked tetraloop structure, wherein the 3′ end of the sense strand comprises a stem-tetraloop structure and comprises two terminal overhang nucleotides at the 3′ end of the antisense strand. In some embodiments, the two terminal overhang nucleotides are GG. Typically, one or both of the two terminal GG nucleotides of the antisense strand are not complementary with the target.

In some embodiments, the 5′ end and/or the 3′end of a sense or antisense strand has an inverted cap nucleotide.

In some embodiments, one or more (e.g., 2, 3, 4, 5, 6) modified internucleotide linkages are provided between terminal nucleotides of the 3′ end or 5′ end of a sense and/or antisense strand. In some embodiments, modified internucleotide linkages are provided between overhang nucleotides at the 3′ end or 5′ end of a sense and/or antisense strand.

Oligonucleotide Modifications

In some embodiments, a dsRNAi oligonucleotide described herein comprises a modification. Oligonucleotides (e.g., dsRNAi oligonucleotides) may be modified in various ways to improve or control specificity, stability, delivery, bioavailability, resistance from nuclease degradation, immunogenicity, base-pairing properties, RNA distribution and cellular uptake and other features relevant to therapeutic or research use.

In some embodiments, the modification is a modified sugar. In some embodiments, the modification is a 5′-terminal phosphate group. In some embodiments, the modification is a modified internucleotide linkage. In some embodiments, the modification is a modified base.

In some embodiments, an oligonucleotide described herein can comprise any one of the modifications described herein or any combination thereof. For example, in some embodiments, an oligonucleotide described herein comprises at least one modified sugar, a 5′-terminal phosphate group, at least one modified internucleotide linkage, and at least one modified base. In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the oligonucleotide comprises at least one modified sugar, a 5′-terminal phosphate group, at least one modified internucleotide linkage, and at least one modified base.

The number of modifications on an oligonucleotide (e.g., a dsRNAi oligonucleotide) and the position of those nucleotide modifications may influence the properties of an oligonucleotide. For example, oligonucleotides may be delivered in vivo by conjugating them to or encompassing them in a lipid nanoparticle (LNP) or similar carrier. However, when an oligonucleotide is not protected by an LNP or similar carrier, it may be advantageous for at least some of the nucleotides to be modified. Accordingly, in some embodiments, all or substantially all the nucleotides of an oligonucleotide are modified. In some embodiments, more than half of the nucleotides are modified. In some embodiments, less than half of the nucleotides are modified. In some embodiments, the sugar moiety of all nucleotides comprising the oligonucleotide is modified at the 2′ position. The modifications may be reversible or irreversible. In some embodiments, an oligonucleotide as disclosed herein has a number and type of modified nucleotides sufficient to cause the desired characteristics (e.g., protection from enzymatic degradation, capacity to target a desired cell after in vivo administration, and/or thermodynamic stability).

Sugar Modifications

In some embodiments, a dsRNAi oligonucleotide described herein comprises a modified sugar. In some embodiments, a modified sugar (also referred herein to a sugar analog) includes a modified deoxyribose or ribose moiety in which, for example, one or more modifications occur at the 2′, 3′, 4′ and/or 5′ carbon position of the sugar. In some embodiments, a modified sugar may also include non-natural alternative carbon structures such as those present in locked nucleic acids (“LNA”; see, e.g., Koshkin et al. (1998) TETRAHEDON 54:3607-30), unlocked nucleic acids (“UNA”; see, e.g., Snead et al. (2013) MOL. THER-NUCL. ACIDS 2:e103) and bridged nucleic acids (“BNA”; see, e.g., Imanishi & Obika (2002) CHEM COMMUN. (CAMB) 21:1653-59).

In some embodiments, a nucleotide modification in a sugar comprises a 2′-modification. In some embodiments, a 2′-modification may be 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-fluoro (2′-F), 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA) or 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA). In some embodiments, the modification is 2′-F, 2′-OMe or 2′-MOE. In some embodiments, a modification in a sugar comprises a modification of the sugar ring, which may comprise modification of one or more carbons of the sugar ring. For example, a modification of a sugar of a nucleotide may comprise a 2′-oxygen of a sugar is linked to a 1′-carbon or 4′-carbon of the sugar, or a 2′-oxygen is linked to the 1′-carbon or 4′-carbon via an ethylene or methylene bridge. In some embodiments, a modified nucleotide has an acyclic sugar that lacks a 2′-carbon to 3′-carbon bond. In some embodiments, a modified nucleotide has a thiol group, e.g., in the 4′ position of the sugar.

In some embodiments, a dsRNAi oligonucleotide described herein comprises at least about 1 modified nucleotide (e.g., at least 1, at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60, or more). In some embodiments, the sense strand of the dsRNAi oligonucleotide comprises at least about 1 modified nucleotide (e.g., at least 1, at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, or more). In some embodiments, the antisense strand of the dsRNAi oligonucleotide comprises at least about 1 modified nucleotide (e.g., at least 1, at least 5, at least 10, at least 15, at least 20, or more).

In some embodiments, all the nucleotides of the sense strand of the dsRNAi oligonucleotide are modified. In some embodiments, all the nucleotides of the antisense strand of the dsRNAi oligonucleotide are modified. In some embodiments, all the nucleotides of the dsRNAi oligonucleotide (i.e., both the sense strand and the antisense strand) are modified. In some embodiments, the modified nucleotide comprises a 2′-modification (e.g., a 2′-F or 2′-OMe, 2′-MOE, and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid).

In some embodiments, the disclosure provides dsRNAi oligonucleotides having different modification patterns. Exemplary modification patterns are set forth in U.S. Provisional Application No. 62/909,278 and in WO 2021/067744, both incorporated herein by this reference. In some embodiments, the modified dsRNAi oligonucleotides comprise a sense strand sequence having a modification pattern as set forth in the Examples and Sequence Listing and an antisense strand having a modification pattern as set forth in the Examples and

SEQUENCE LISTING

In some embodiments, a dsRNAi oligonucleotide disclosed herein comprises an antisense strand having nucleotides that are modified with 2′-F. In some embodiments, a dsRNAi oligonucleotide disclosed herein comprises an antisense strand comprising nucleotides that are modified with 2′-F and 2′-OMe. In some embodiments, a dsRNAi oligonucleotide disclosed herein comprises a sense strand having nucleotides that are modified with 2′-F. In some embodiments, a dsRNAi oligonucleotide disclosed herein comprises a sense strand comprising nucleotides that are modified with 2′-F and 2′-OMe.

In some embodiments, a dsRNAi oligonucleotide described herein comprises a sense strand with about 10-15%, 10%, 11%, 12%, 13%, 14% or 15% of the nucleotides of the sense strand comprising a 2′-fluoro modification. In some embodiments, about 11% of the nucleotides of the sense strand comprise a 2-fluoro modification. In some embodiments, a dsRNAi oligonucleotide described herein comprises an antisense strand with about 25-35%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34% or 35% of the nucleotides of the antisense strand comprising a 2′-fluoro modification. In some embodiments, about 32% of the nucleotides of the antisense strand comprise a 2′-fluoro modification. In some embodiments, the dsRNAi oligonucleotide has about 15-25%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, or 25% of its nucleotides comprising a 2′-fluoro modification. In some embodiments, about 19% of the nucleotides in the dsRNAi oligonucleotide comprise a 2′-fluoro modification.

In some embodiments, one or more of positions 8, 9, 10 or 11 of the sense strand is modified with a 2′-F group. In some embodiments, one or more of positions 3, 8, 9, 10, 12, 13 and 17 of the sense strand is modified with a 2′-F group. In some embodiments, one or more of positions 2, 3, 4, 5, 7, 10 and 14 of the antisense strand is modified with a 2′-F group. In some embodiments, one or more of positions 2, 3, 4, 5, 7, 8, 10, 14, 16 and 19 of the antisense strand is modified with a 2′-F group. In some embodiments, the sugar moiety at each of nucleotides at positions 1-7 and 12-20 in the sense strand is modified with a 2′-OMe. In some embodiments, the sugar moiety at each of nucleotides at positions 1-7, 12-27 and 31-36 in the sense strand is modified with a 2′-OMe. In some embodiments, the sugar moiety at each of nucleotides at positions 1-2, 4-7, 11, 14-16 and 18-20 in the sense strand is modified with a 2′-OMe. In some embodiments, the sugar moiety at each of nucleotides at positions 1-2, 4-7, 11, 14-16, 18-27 and 31-36 in the sense strand is modified with a 2′-OMe. In some embodiments, the sugar moiety at each of nucleotides at positions 1, 6, 8-9, 11-13, and 15-22 in the antisense strand is modified with a 2′-OMe. In some embodiments, the sugar moiety at each of nucleotides at positions 6, 9, 11-13, 15, 17, 18 and 20-22 in the antisense strand is modified with a 2′-OMe. In some embodiments, the sugar moiety at each of nucleotides at positions 1, 6, 9, 11-13, 15, 17, 18 and 20-22 in the antisense strand is modified with a 2′-OMe.

In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein one or more of positions 8, 9, 10 or 11 of the sense strand is modified with a 2′-F group.

In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein one or more of positions 3, 8, 9, 10, 12, 13 and 17 of the sense strand is modified with a 2′-F group.

In some embodiments, the antisense strand has 3 nucleotides that are modified at the 2′-position of the sugar moiety with a 2′-F. In some embodiments, the sugar moiety at positions 2, 5 and 14 and optionally up to 3 of the nucleotides at positions 1, 3, 7 and 10 of the antisense strand are modified with a 2′-F. In other embodiments, the sugar moiety at each of the positions 2, 5 and 14 of the antisense strand is modified with the 2′-F. In other embodiments, the sugar moiety at each of the positions 1, 2, 5 and 14 of the antisense strand is modified with the 2′-F. In still other embodiments, the sugar moiety at each of the positions 1, 2, 3, 5, 7 and 14 of the antisense strand is modified with the 2′-F. In yet another embodiment, the sugar moiety at each of the positions 1, 2, 3, 5, 10 and 14 of the antisense strand is modified with the 2′-F. In another embodiment, the sugar moiety at each of the positions 2, 3, 5, 7, 10 and 14 of the antisense strand is modified with the 2′-F.

In some embodiments, the antisense strand has 3 nucleotides that are modified at the 2′-position of the sugar moiety with a 2′-F. In some embodiments, the sugar moiety at positions 2, 5 and 14 and optionally up to 3 of the nucleotides at positions 3, 4, 7 and 10 of the antisense strand are modified with a 2′-F. In other embodiments, the sugar moiety at each of positions 2, 5 and 14 of the antisense strand is modified with the 2′-F. In other embodiments, the sugar moiety at each of positions 2, 4, 5 and 14 of the antisense strand is modified with the 2′-F. In still other embodiments, the sugar moiety at each of positions 2, 3, 4, 5, 7 and 14 of the antisense strand is modified with the 2′-F. In yet another embodiment, the sugar moiety at each of positions 2, 3, 4, 5, 10 and 14 of the antisense strand is modified with the 2′-F. In another embodiment, the sugar moiety at each of positions 2, 3, 4, 5, 7, 10 and 14 of the antisense strand is modified with the 2′-F. In some embodiments, the sugar moiety at each of positions 2, 3, 4, 5, 7, 8, 10, 14, 16 and 19 is modified with the 2′-F.

In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the sugar moiety at one or more positions 2, 3, 4, 5, 7, 10 and 14 of the antisense strand is modified with the 2′-F.

In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the sugar moiety at one or more positions 2, 3, 4, 5, 7, 8, 10, 14, 16 and 19 of the antisense strand is modified with the 2′-F.

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 2 and 14 modified with 2′-F. In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 2, 5, and 14 modified with 2′-F. In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 1, 2, 5, and 14 modified with 2′-F. In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 1, 2, 3, 5, 7, and 14 modified with 2′-F.

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 1, 2, 3, 5, 10, and 14 modified with 2′-F.

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 2 and 14 modified with 2′-F. In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 2, 5, and 14 modified with 2′-F. In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 2, 4, 5, and 14 modified with 2′-F. In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 2, 3, 4, 5, 7, and 14 modified with 2′-F. In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 2, 3, 4, 5, 10, and 14 modified with 2′-F. In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 2, 3, 4, 5, 7, 10 and 14 modified with 2′-F. In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at positions 2, 3, 4, 5, 7, 8, 10, 14, 16 and 19 modified with 2′-F.

In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein one or more of positions 8, 9, 10 or 11 of the sense strand and one or more positions 2, 3, 4, 5, 7, 10 and 14 of the antisense strand is modified with the 2′-F.

In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein one or more of positions 3, 8, 9, 10, 12, 13 and 17 of the sense strand and one or more positions 2, 3, 4, 5, 7, 8, 10, 14, 16 and 19 of the antisense strand is modified with the 2′-F.

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety of each of the nucleotides at positions 2, 5, and 14 of the antisense strand modified with 2′-F and the sugar moiety of each of the remaining nucleotides of the antisense strand modified with a modification selected from the group consisting of 2′-0-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-0-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety of each of the nucleotides at positions 1, 2, 5, and 14 of the antisense strand modified with 2′-F and the sugar moiety of each of the remaining nucleotides of the antisense strand modified with a modification selected from the group consisting of 2′-0-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-0-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety of each of the nucleotides at positions 2, 4, 5, and 14 of the antisense strand modified with 2′-F and the sugar moiety of each of the remaining nucleotides of the antisense strand modified with a modification selected from the group consisting of 2′-0-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-0-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety of each of the nucleotides at positions 1, 2, 3, 5, 7, and 14 of the antisense strand modified with 2′-F and the sugar moiety of each of the remaining nucleotides of the antisense strand modified with a modification selected from the group consisting of 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety of each of the nucleotides at positions 2, 3, 4, 5, 7, and 14 of the antisense strand modified with 2′-F and the sugar moiety of each of the remaining nucleotides of the antisense strand modified with a modification selected from the group consisting of 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety of each of the nucleotides at positions 1, 2, 3, 5, 10, and 14 of the antisense strand modified with 2′-F and the sugar moiety of each of the remaining nucleotides of the antisense strand modified with a modification selected from the group consisting of 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety of each of the nucleotides at positions 2, 3, 4, 5, 10, and 14 of the antisense strand modified with 2′-F and the sugar moiety of each of the remaining nucleotides of the antisense strand modified with a modification selected from the group consisting of 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety of each of the nucleotides at positions 2, 3, 5, 7, 10, and 14 of the antisense strand modified with 2′-F and the sugar moiety of each of the remaining nucleotides of the antisense strand modified with a modification selected from the group consisting of 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety of each of the nucleotides at positions 2, 3, 4, 5, 7, 10, and 14 of the antisense strand modified with 2′-F and the sugar moiety of each of the remaining nucleotides of the antisense strand modified with a modification selected from the group consisting of 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety of each of the nucleotides at positions 2, 3, 4, 5, 7, 8, 10, 14, 16 and 19 of the antisense strand modified with 2′-F and the sugar moiety of each of the remaining nucleotides of the antisense strand modified with a modification selected from the group consisting of 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8, position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, position 21, or position 22 modified with 2′-F.

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8, position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, position 21, or position 22 modified with 2′-OMe.

In some embodiments, an oligonucleotide provided herein comprises an antisense strand having the sugar moiety at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8, position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, position 21, or position 22 modified with a modification selected from the group consisting of 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises a sense strand having the sugar moiety at positions 8-11 modified with 2′-F. In some embodiments, an oligonucleotide provided herein comprises a sense strand having the sugar moiety at positions 3, 8, 9, 10, 12, 13 and 17 modified with 2′-F. In some embodiments, an oligonucleotide provided herein comprises a sense strand having the sugar moiety at positions 1-7 and 12-17 or 12-20 modified with 2′OMe. In some embodiments, an oligonucleotide provided herein comprises a sense strand having the sugar moiety of each of the nucleotides at positions 1-7 and 12-17 or 12-20 of the sense strand modified with a modification selected from the group consisting of 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA). In some embodiments, an oligonucleotide provided herein comprises a sense strand having the sugar moiety at positions 1-2, 4-7, 11, 14-16 and 18-20 modified with 2′OMe. In some embodiments, an oligonucleotide provided herein comprises a sense strand having the sugar moiety of each of the nucleotides at positions 1-2, 4-7, 11, 14-16 and 18-20 of the sense strand modified with a modification selected from the group consisting of 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

In some embodiments, an oligonucleotide provided herein comprises a sense strand having the sugar moiety at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8, position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, position 21, position 22, position 23, position 24, position 25, position 26, position 27, position 28, position 29, position 30, position 31, position 32, position 33, position 34, position 35, or position 36 modified with 2′-F.

In some embodiments, an oligonucleotide provided herein comprises a sense strand having the sugar moiety at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8, position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, position 21, position 22, position 23, position 24, position 25, position 26, position 27, position 28, position 29, position 30, position 31, position 32, position 33, position 34, position 35, or position 36 modified with 2′-OMe.

In some embodiments, an oligonucleotide provided herein comprises a sense strand having the sugar moiety at position 1, position 2, position 3, position 4, position 5, position 6, position 7, position 8, position 9, position 10, position 11, position 12, position 13, position 14, position 15, position 16, position 17, position 18, position 19, position 20, position 21, position 22, position 23, position 24, position 25, position 26, position 27, position 28, position 29, position 30, position 31, position 32, position 33, position 34, position 35, or position 36 modified with a modification selected from the group consisting of 2′-O-propargyl, 2′-O-propylamin, 2′-amino, 2′-ethyl, 2′-aminoethyl (EA), 2′-O-methyl (2′-OMe), 2′-O-methoxyethyl (2′-MOE), 2′-O-[2-(methylamino)-2-oxoethyl] (2′-O-NMA), and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′-FANA).

5′-Terminal Phosphate

In some embodiments, an oligonucleotide described herein comprises a 5′-terminal phosphate. In some embodiments, 5′-terminal phosphate groups of an RNAi oligonucleotide enhance the interaction with Ago2. However, oligonucleotides comprising a 5′-phosphate group may be susceptible to degradation via phosphatases or other enzymes, which can limit their bioavailability in vivo. In some embodiments, an oligonucleotide (e.g., a double-stranded oligonucleotide) herein includes analogs of 5′ phosphates that are resistant to such degradation. In some embodiments, the phosphate analog is oxymethylphosphonate, vinylphosphonate or malonylphosphonate, or a combination thereof. In certain embodiments, the 5′ end of an oligonucleotide strand is attached to chemical moiety that mimics the electrostatic and steric properties of a natural 5′-phosphate group (“phosphate mimic”). In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the oligonucleotide comprises a 5′-terminal phosphate.

In some embodiments, an oligonucleotide has a phosphate analog at a 4′-carbon position of the sugar (referred to as a “4′-phosphate analog”). See, e.g., Intl. Patent Application Publication No. WO 2018/045317. In some embodiments, an oligonucleotide herein comprises a 4′-phosphate analog at a 5′-terminal nucleotide. In some embodiments, a phosphate analog is an oxymethylphosphonate, in which the oxygen atom of the oxymethyl group is bound to the sugar moiety (e.g., at its 4′-carbon) or analog thereof. In other embodiments, a 4′-phosphate analog is a thiomethylphosphonate or an aminomethylphosphonate, in which the sulfur atom of the thiomethyl group or the nitrogen atom of the amino methyl group is bound to the 4′-carbon of the sugar moiety or analog thereof. In certain embodiments, a 4′-phosphate analog is an oxymethylphosphonate. In some embodiments, an oxymethylphosphonate is represented by the formula —O—CH₂—PO(OH)₂, —O—CH₂—PO(OR)₂, or —O—CH₂—POOH(R), in which R is independently selected from H, CH₃, an alkyl group, CH₂CH₂CN, CH₂OCOC(CH₃)₃, CH₂OCH₂CH₂Si (CH₃)₃ or a protecting group. In certain embodiments, the alkyl group is CH₂CH₃. More typically, R is independently selected from H, CH₃ or CH₂CH₃. In some embodiment, R is CH₃. In some embodiments, the 4′-phosphate analog is 5′-methoxyphosphonate-4′-oxy.

In some embodiments, a dsRNAi oligonucleotide provided herein comprises an antisense strand comprising a 4′-phosphate analog at the 5′-terminal nucleotide, wherein 5′-terminal nucleotide comprises the following structure:

5′-methoxyphosphonate-4′-oxy-2′-O-methyluridine phosphorothioate [MePhosphonate-4O-mUs]

Modified Internucleotide Linkage

In some embodiments, an oligonucleotide (e.g., a dsRNAi oligonucleotide) herein comprises a modified internucleotide linkage. In some embodiments, phosphate modifications or substitutions result in an oligonucleotide that comprises at least about 1 (e.g., at least 1, at least 2, at least 3 or at least 5) modified internucleotide linkage. In some embodiments, any one of the oligonucleotides disclosed herein comprises about 1 to about 10 (e.g., 1 to 10, 2 to 8, 4 to 6, 3 to 10, 5 to 10, 1 to 5, 1 to 3 or 1 to 2) modified internucleotide linkages. In some embodiments, any one of the oligonucleotides disclosed herein comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 modified internucleotide linkages.

A modified internucleotide linkage may be a phosphorodithioate linkage, a phosphorothioate linkage, a phosphotriester linkage, a thionoalkylphosphonate linkage, a thionalkylphosphotriester linkage, a phosphoramidite linkage, a phosphonate linkage or a boranophosphate linkage. In some embodiments, at least one modified internucleotide linkage of any one of the oligonucleotides as disclosed herein is a phosphorothioate linkage.

In some embodiments, an oligonucleotide provided herein (e.g., a dsRNAi oligonucleotide) has a phosphorothioate linkage between one or more of positions 1 and 2 of the sense strand, positions 1 and 2 of the antisense strand, positions 2 and 3 of the antisense strand, positions 3 and 4 of the antisense strand, positions 20 and 21 of the antisense strand, and positions 21 and 22 of the antisense strand. In some embodiments, the oligonucleotide described herein has a phosphorothioate linkage between each of positions 1 and 2 of the sense strand, positions 1 and 2 of the antisense strand, positions 2 and 3 of the antisense strand, positions 20 and 21 of the antisense strand, and positions 21 and 22 of the antisense strand. In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the oligonucleotide comprises a modified internucleotide linkage. In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:
  • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and,
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the oligonucleotide comprises a phosphorothioate linkage between one or more of positions 1 and 2 of the sense strand, positions 1 and 2 of the antisense strand, positions 2 and 3 of the antisense strand, positions 3 and 4 of the antisense strand, positions 20 and 21 of the antisense strand, and positions 21 and 22 of the antisense strand. In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:
  • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the oligonucleotide comprises a phosphorothioate linkage between each of positions 1 and 2 of the sense strand, positions 1 and 2 of the antisense strand, positions 2 and 3 of the antisense strand, positions 20 and 21 of the antisense strand, and positions 21 and 22 of the antisense strand.

Base Modifications

In some embodiments, oligonucleotides herein (e.g., dsRNAi oligonucleotides) have one or more modified nucleobases. In some embodiments, modified nucleobases (also referred to herein as base analogs) are linked at the 1′ position of a nucleotide sugar moiety. In certain embodiments, a modified nucleobase is a nitrogenous base. In certain embodiments, a modified nucleobase does not contain nitrogen atom. See, e.g., US Patent Application Publication No. 2008/0274462. In some embodiments, a modified nucleotide comprises a universal base. In some embodiments, a modified nucleotide does not contain a nucleobase (abasic). In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the oligonucleotide comprises one or more modified nucleobases.

In some embodiments, a universal base is a heterocyclic moiety located at the 1′ position of a nucleotide sugar moiety in a modified nucleotide, or the equivalent position in a nucleotide sugar moiety substitution, that, when present in a duplex, can be positioned opposite more than one type of base without substantially altering structure of the duplex. In some embodiments, compared to a reference single-stranded nucleic acid (e.g., oligonucleotide) that is fully complementary to a target nucleic acid, a single-stranded nucleic acid containing a universal base forms a duplex with the target nucleic acid that has a lower Tri, than a duplex formed with the complementary nucleic acid. In some embodiments, when compared to a reference single-stranded nucleic acid in which the universal base has been replaced with a base to generate a single mismatch, the single-stranded nucleic acid containing the universal base forms a duplex with the target nucleic acid that has a higher T_m than a duplex formed with the nucleic acid comprising the mismatched base.

Non-limiting examples of universal-binding nucleotides include, but are not limited to, inosine, 1-β-D-ribofuranosyl-5-nitroindole and/or 1-β-D-ribofuranosyl-3-nitropyrrole (see, US Patent Application Publication No. 2007/0254362; Van Aerschot et al. (1995) NUCLEIC ACIDS RES. 23:4363-4370; Loakes et al. (1995) NUCLEIC ACIDS RES. 23:2361-66; and Loakes & Brown (1994) NUCLEIC ACIDS RES. 22:4039-43).

Targeting Ligands

In some embodiments, it is desirable to target the oligonucleotides of the disclosure (e.g., dsRNAi oligonucleotides) to one or more cells or one or more organs. Such a strategy can help to avoid undesirable effects in other organs or avoid undue loss of the oligonucleotide to cells, tissue or organs that would not benefit from the oligonucleotide. Accordingly, in some embodiments, oligonucleotides disclosed herein (e.g., dsRNAi oligonucleotides) are modified to facilitate targeting and/or delivery to a particular tissue, cell, or organ (e.g., to facilitate delivery of the oligonucleotide to the liver). In some embodiments, an oligonucleotide comprises at least one nucleotide (e.g., 1, 2, 3, 4, 5, 6 or more nucleotides) conjugated to one or more targeting ligand(s). In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the oligonucleotide comprises a targeting ligand conjugated to at least once nucleotide,

In some embodiments, the targeting ligand comprises a carbohydrate, amino sugar, cholesterol, peptide, polypeptide, protein, or part of a protein (e.g., an antibody or antibody fragment), or lipid. In some embodiments, the targeting ligand is an aptamer. For example, a targeting ligand may be an RGD peptide that is used to target tumor vasculature or glioma cells, CREKA peptide to target tumor vasculature or stoma, transferring, lactoferrin, or an aptamer to target transferrin receptors expressed on CNS vasculature, or an anti-EGFR antibody to target EGFR on glioma cells. In certain embodiments, the targeting ligand is one or more GalNAc moieties.

In some embodiments, 1 or more (e.g., 1, 2, 3, 4, 5 or 6) nucleotides of an oligonucleotide are each conjugated to a separate targeting ligand. In some embodiments, 2 to 4 nucleotides of an oligonucleotide are each conjugated to a separate targeting ligand. In some embodiments, targeting ligands are conjugated to 2 to 4 nucleotides at either ends of the sense or antisense strand (e.g., targeting ligands are conjugated to a 2 to 4 nucleotide overhang or extension on the 5′ or 3′ end of the sense or antisense strand) such that the targeting ligands resemble bristles of a toothbrush and the oligonucleotide resembles a toothbrush. For example, an oligonucleotide may comprise a stem-loop at either the 5′ or 3′ end of the sense strand and 1, 2, 3 or 4 nucleotides of the loop of the stem may be individually conjugated to a targeting ligand. In some embodiments, an oligonucleotide (e.g., a dsRNAi oligonucleotide) provided by the disclosure comprises a stem-loop at the 3′ end of the sense strand, wherein the loop of the stem-loop comprises a triloop or a tetraloop, and wherein the 3 or 4 nucleotides comprising the triloop or tetraloop, respectively, are individually conjugated to a targeting ligand.

GalNAc is a high affinity ligand for the ASGPR, which is primarily expressed on the sinusoidal surface of hepatocyte cells and has a major role in binding, internalizing and subsequent clearing circulating glycoproteins that contain terminal galactose or GalNAc residues (asialoglycoproteins). Conjugation (either indirect or direct) of GalNAc moieties to oligonucleotides of the instant disclosure can be used to target these oligonucleotides to the ASGPR expressed on cells. In some embodiments, an oligonucleotide of the instant disclosure is conjugated to at least one or more GalNAc moieties, wherein the GalNAc moieties target the oligonucleotide to an ASGPR expressed on human liver cells (e.g., human hepatocytes). In some embodiments, the GalNAc moiety targets the oligonucleotide to the liver.

In some embodiments, an oligonucleotide of the instant disclosure is conjugated directly or indirectly to a monovalent GalNAc. In some embodiments, the oligonucleotide is conjugated directly or indirectly to more than one monovalent GalNAc (i.e., is conjugated to 2, 3 or 4 monovalent GalNAc moieties, and is typically conjugated to 3 or 4 monovalent GalNAc moieties). In some embodiments, an oligonucleotide is conjugated to one or more bivalent GalNAc, trivalent GalNAc or tetravalent GalNAc moieties.

In some embodiments, 1 or more (e.g., 1, 2, 3, 4, 5 or 6) nucleotides of an oligonucleotide are each conjugated to a GalNAc moiety. In some embodiments, 2 to 4 nucleotides of a tetraloop are each conjugated to a separate GalNAc. In some embodiments, 1 to 3 nucleotides of a triloop are each conjugated to a separate GalNAc. In some embodiments, targeting ligands are conjugated to 2 to 4 nucleotides at either ends of the sense or antisense strand (e.g., ligands are conjugated to a 2 to 4 nucleotide overhang or extension on the 5′ or 3′ end of the sense or antisense strand) such that the GalNAc moieties resemble bristles of a toothbrush and the oligonucleotide resembles a toothbrush. In some embodiments, GalNAc moieties are conjugated to a nucleotide of the sense strand. For example, three (3) or four (4) GalNAc moieties can be conjugated to nucleotides in the tetraloop of the sense strand where each GalNAc moiety is conjugated to 1 nucleotide.

In some embodiments, the tetraloop is any combination of adenine and guanine nucleotides.

In some embodiments, the tetraloop (L) has a monovalent GalNAc moiety attached to any one or more guanine nucleotides of the tetraloop via any linker described herein, as depicted below (X=heteroatom):

In some embodiments, the tetraloop (L) has a monovalent GalNAc attached to any one or more adenine nucleotides of the tetraloop via any linker described herein, as depicted below (X=heteroatom):

In some embodiments, an oligonucleotide herein comprises a monovalent GalNAc attached to a guanine nucleotide referred to as [ademG-GalNAc] or 2′-aminodiethoxymethanol-Guanine-GalNAc, as depicted below:

In some embodiments, an oligonucleotide herein comprises a monovalent GalNAc attached to an adenine nucleotide, referred to as [ademA-GalNAc] or 2′-aminodiethoxymethanol-Adenine-GalNAc, as depicted below:

An example of such conjugation is shown below for a loop comprising from 5′ to 3′ the nucleotide sequence GAAA (L=linker, X=heteroatom). Such a loop may be present, for example, at positions 27-30 of a sense strand provided herein, as shown in FIGS. 2 and 4A. In the chemical formula,

is used to describe an attachment point to the oligonucleotide strand.

Appropriate methods or chemistry (e.g., click chemistry) can be used to link a targeting ligand to a nucleotide. In some embodiments, a targeting ligand is conjugated to a nucleotide using a click linker. In some embodiments, an acetal-based linker is used to conjugate a targeting ligand to a nucleotide of any one of the oligonucleotides described herein. Acetal-based linkers are disclosed, for example, in Intl. Patent Application Publication No. WO 2016/100401. In some embodiments, the linker is a labile linker. However, in other embodiments, the linker is stable. Examples are shown below for a loop comprising from 5′ to 3′ the nucleotides GAAA, in which GalNAc moieties are attached to nucleotides of the loop using an acetal linker. Such a loop may be present, for example, at positions 27-30 of the sense strand as shown in FIGS. 2 and 4A. In the chemical formula,

is an attachment point to the oligonucleotide strand.

As mentioned, various appropriate methods or chemistry synthetic techniques (e.g., click chemistry) can be used to link a targeting ligand to a nucleotide. In some embodiments, a targeting ligand is conjugated to a nucleotide using a click linker. In some embodiments, an acetal-based linker is used to conjugate a targeting ligand to a nucleotide of any one of the oligonucleotides described herein. Acetal-based linkers are disclosed, for example, in Intl. Patent Application Publication No. WO 2016/100401. In some embodiments, the linker is a labile linker. However, in other embodiments, the linker is a stable linker.

In some embodiments, a duplex extension (e.g., of up to 3, 4, 5 or 6 bp in length) is provided between a targeting ligand (e.g., a GalNAc moiety) and a dsRNA. In some embodiments, the oligonucleotides herein (e.g., dsRNAi oligonucleotides) do not have a GalNAc conjugated thereto.

In some embodiments, the sense and antisense strands of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the oligonucleotide comprises at least one GalNAc moiety conjugated to a nucleotide.Exemplary KHK-Targeting dsRNAi Oligonucleotides

In some embodiments, the disclosure provides dsRNAi oligonucleotides that target KHK mRNA and reduce KHK expression (referred to herein as KHK-targeting dsRNAi oligonucleotides), wherein the oligonucleotides comprise a sense strand and an antisense strand that form a duplex region, and wherein the antisense strand comprises a region of complementarity to KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is at least 15 contiguous nucleotides in length. In some embodiments, the disclosure provides dsRNAi oligonucleotides that target KHK mRNA and reduce KHK expression (referred to herein as KHK-targeting dsRNAi oligonucleotides), wherein the oligonucleotides comprise a sense strand and an antisense strand that form a duplex region, and wherein the antisense strand comprises a region of complementarity to KHK mRNA target sequence of nucleotides 1-19 of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is at least 15 contiguous nucleotides in length. In some embodiments, the region of complementarity is 15-20 nucleotides in length. In some embodiments, the region of complementarity is 15 nucleotides, 16 nucleotides, 17 nucleotides, 18 nucleotides, 19 nucleotides, or 20 nucleotides in length. In some embodiments, the region of complementarity is at least 19 contiguous nucleotides in length. In some embodiments, the region of complementarity is at least 20 nucleotides in length. In some embodiments, the region of complementarity is 19 nucleotides in length. In some embodiments, the region of complementarity is 20 nucleotides in length.

In some embodiments, the sense strand is 15 to 50 nucleotides in length. In some embodiments, the sense strand is 18 to 36 nucleotides in length. In some embodiments, the sense strand comprises a nucleotide sequence selected from SEQ ID NOs: 909, 894, 897, 892, 891 and 887, and is 15 to 50 nucleotides in length. In some embodiments, the sense strand is 36 nucleotides in length. In some embodiments, the antisense strand is 15 to 30 nucleotides in length. In some embodiments, the antisense strand comprises a nucleotide sequence selected from SEQ ID NOs: 936, 920, 923, 917, 918 and 913, and is 15 to 50 nucleotides in length. In some embodiments, the antisense strand is 22 nucleotides in length. In some embodiments, the sense strand is 36 nucleotides in length and the antisense strand is 22 nucleotides in length and the sense and antisense strand form a duplex region that is at least 19 nucleotides in length. In some embodiments, the duplex region is 20 nucleotides in length.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides for reducing KHK expression provided by the disclosure comprise a stem-loop set forth as S1-L-S2, wherein S1 is complementary to S2, and wherein L forms a loop between S1 and S2 of 3-5 nucleotides in length. In some embodiments, 51 and S2 are 1-10 nucleotides in length and are the same length. In some embodiments, 51 and S2 are 1 nucleotide, 2 nucleotides, 3 nucleotides, 4 nucleotides, 5 nucleotides, 6 nucleotides, 7 nucleotides, 8 nucleotides, 9 nucleotides, or 10 nucleotides in length. In some embodiments, 51 and S2 are 6 nucleotides in length. In some embodiments the loop is 3 nucleotides in length. In some embodiments, the loop is 4 nucleotides in length. In some embodiments, the loop is 5 nucleotides in length. In some embodiments, L is a triloop or a tetraloop. In some embodiments, L is a triloop. In some embodiments, L is a tetraloop. In some embodiments, the tetraloop comprises the sequence 5′-GAAA-3′. In some embodiments, the stem loop comprises the sequence 5′-GCAGCCGAAAGGCUGC-3′ (SEQ ID NO: 871). In some embodiments, up to 4 nucleotides comprising L are each conjugated to a targeting ligand. In some embodiments, 1 nucleotide, 2 nucleotides, 3 nucleotides, or 4 nucleotides comprising L are each conjugated to a targeting ligand. In some embodiments, 3 nucleotides comprising L are each conjugated to a targeting ligand. In some embodiments, L is a tetraloop comprising the sequence 5′-GAAA-3′, wherein each adenosine (A) nucleoside comprising the tetraloop is conjugated to a targeting ligand comprising a monovalent N-acetylgalactosamine (GalNAc) moiety.

In some embodiments, the antisense strand comprises a 3′ overhang of one or more nucleotides in length. In some embodiments, the 3′ overhang is two (2) nucleotides in length. In some embodiments, the sequence of the 3′ overhang is 5′-GG-3′.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides for reducing KHK expression provided by the disclosure comprise a sense strand of 36 nucleotides in length and an antisense strand of 22 nucleotides in length, wherein the sense strand and the antisense strand form a duplex region of at least 19 nucleotides in length, optionally 20 nucleotides in length, wherein the 3′ end of the sense strand comprises a stem-loop set forth as S1-L-S2, wherein S1 is complementary to S2, and wherein L forms a loop between S1 and S2 of 3-5 nucleotides in length, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is 19 contiguous nucleotides in length, optionally 20 nucleotides in length. In some embodiments, the KHK-targeting dsRNAi oligonucleotides for reducing KHK expression provided by the disclosure comprise a sense strand of 36 nucleotides in length and an antisense strand of 22 nucleotides in length, wherein the sense strand and the antisense strand form a duplex region of at least 19 nucleotides in length, optionally 20 nucleotides in length, wherein the 3′ end of the sense strand comprises a stem-loop set forth as S1-L-S2, wherein S1 is complementary to S2, and wherein L forms a loop between S1 and S2 of 3-5 nucleotides in length, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of nucleotides 1-19 of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is 19 contiguous nucleotides in length, optionally 20 nucleotides in length.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides for reducing KHK expression provided by the disclosure comprises at least one modified nucleotide. In some embodiments, the modified nucleotide comprises a five (5) carbon sugar (e.g., ribose) with a 2′-modification. In some embodiments, the 2′-modification is a modification selected from 2′-aminoethyl, 2′-fluoro, 2′-O-methyl, 2′-O-methoxyethyl, and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid. In some embodiments, the 2′-modification is 2′-fluoro or 2′-O-methyl. In some embodiments, all nucleotides comprising the KHK-targeting dsRNAi oligonucleotides are modified. In some embodiments, all nucleotides comprising the KHK-targeting dsRNAi oligonucleotides are modified with a 2′-modification selected from 2′-fluoro and 2′-O-methyl. In some embodiments, all nucleotides comprising the KHK-targeting dsRNAi oligonucleotides are modified with a combination of 2′-fluoro and 2′-O-methyl. In some embodiments, the sense and antisense strand of an oligonucleotide comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 909 and 936, respectively;
  • (b) SEQ ID NOs: 894 and 920, respectively;
  • (c) SEQ ID NOs: 897 and 923, respectively;
  • (d) SEQ ID NOs: 892 and 918, respectively;
  • (e) SEQ ID NOs: 891 and 917, respectively; and
  • (f) SEQ ID NOs: 887 and 913, respectively;wherein the oligonucleotide is modified with a combination of 2′-fluoro and 2′-O-methyl.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides comprises at least one modified internucleotide linkage. In some embodiments, the at least one modified internucleotide linkage is a phosphorothioate linkage.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides comprise an antisense strand wherein the 4′-carbon of the sugar of the 5′-terminal nucleotide of the antisense strand comprises a phosphate analog. In some embodiments, the phosphate analog is oxymethylphosphonate, vinylphosphonate or malonylphosphonate. In some embodiments, the phosphate analog is a 4′-phosphate analog comprising 5′-methoxyphosphonate-4′-oxy.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides for reducing KHK expression provided by the disclosure comprise a sense strand and an antisense strand, wherein all nucleotides comprising the sense strand and antisense strand are modified, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is at least 15 contiguous nucleotides in length. In some embodiments, the KHK-targeting dsRNAi oligonucleotides for reducing KHK expression provided by the disclosure comprise a sense strand and an antisense strand, wherein all nucleotides comprising the sense strand and antisense strand are modified, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of nucleotides 1-19 of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is at least 15 contiguous nucleotides in length. In some embodiments, the 5′-terminal nucleotide of the antisense strand comprises 5′-methoxyphosphonate-4′-oxy-2′-O-methyluridine [MePhosphonate-4O-mU], as described herein. In some embodiments, the 5′-terminal nucleotide of the antisense strand comprises a phosphorothioate linkage. In some embodiments, the antisense strand and the sense strand comprise one or more 2′-fluoro (2′-F) and 2′-O-methyl (2′-OMe) modified nucleotides and at least one phosphorothioate linkage. In some embodiments, the antisense strand comprises four (4) phosphorothioate linkages and the sense strand comprises one (1) phosphorothioate linkage. In some embodiments, the antisense strand comprises five (5) phosphorothioate linkages and the sense strand comprises one (1) phosphorothioate linkage.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides for reducing KHK expression comprise:

a sense strand comprising a 2′-F modified nucleotide at positions 8-11, a 2′-OMe modified nucleotide at positions 1-7, 12-27, and 31-36, a GalNAc-conjugated nucleotide at position 28, 29 and 30; and a phosphorothioate linkage between positions 1 and 2;

an antisense strand comprising a 2′-F modified nucleotide at positions 2, 3, 4, 5, 7, 10 and 14, a 2′-OMe at positions 1, 6, 8, 9, 11-13, and 15-22, a phosphorothioate linkage between positions 1 and 2, positions 2 and 3, positions 3 and 4, positions 20 and 21, and positions 21 and 22, and a 5′-terminal nucleotide at position 1 comprising a 4′-phosphate analog, optionally wherein the 5′-terminal nucleotide comprises 5′-methoxyphosphonate-4′-oxy-2′-O-methyluridine [MePhosphonate-4O-mU]; wherein positions 1-20 of the antisense strand form a duplex region with positions 1-20 of the sense strand, wherein positions 21-36 of the sense strand form a stem-loop, wherein positions 27-30 form the loop of the stem-loop, optionally wherein positions 27-30 comprise a tetraloop, wherein positions 21 and 22 of the antisense strand comprise an overhang, and wherein the sense strand and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 886 and 912, respectively;
  • (b) SEQ ID NOs: 887 and 913, respectively;
  • (c) SEQ ID NOs: 910 and 937, respectively;
  • (d) SEQ ID NOs: 888 and 914, respectively;
  • (e) SEQ ID NOs: 889 and 915, respectively;
  • (f) SEQ ID NOs: 890 and 916, respectively;
  • (g) SEQ ID NOs: 891 and 917, respectively;
  • (h) SEQ ID NOs: 877 and 884, respectively;
  • (i) SEQ ID NOs: 878 and 930, respectively;
  • (j) SEQ ID NOs: 876 and 883, respectively;
  • (k) SEQ ID NOs: 875 and 882, respectively;
  • (l) SEQ ID NOs: 892 and 918, respectively;
  • (m) SEQ ID NOs: 893 and 919, respectively;
  • (n) SEQ ID NOs: 894 and 920, respectively;
  • (o) SEQ ID NOs: 904 and 931, respectively;
  • (p) SEQ ID NOs: 895 and 921, respectively;
  • (q) SEQ ID NOs: 905 and 932, respectively;
  • (r) SEQ ID NOs: 896 and 922, respectively;
  • (s) SEQ ID NOs: 911 and 938, respectively;
  • (t) SEQ ID NOs: 906 and 933, respectively;
  • (u) SEQ ID NOs: 897 and 923, respectively;
  • (v) SEQ ID NOs: 907 and 934, respectively;
  • (w) SEQ ID NOs: 908 and 935, respectively;
  • (x) SEQ ID NOs: 903 and 929, respectively;
  • (y) SEQ ID NOs: 901 and 927, respectively;
  • (z) SEQ ID NOs: 874 and 881, respectively;
  • (aa) SEQ ID NOs: 902 and 928, respectively;
  • (bb) SEQ ID NOs: 873 and 880, respectively;
  • (cc) SEQ ID NOs: 872 and 879, respectively;
  • (dd) SEQ ID NOs: 898 and 924, respectively;
  • (ee) SEQ ID NOs: 899 and 925, respectively
  • (ff) SEQ ID NOs: 900 and 926, respectively; and
  • (gg) SEQ ID NOs: 909 and 936, respectively.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides for reducing KHK expression comprise:

a sense strand comprising a 2′-F modified nucleotide at positions 8-11, a 2′-OMe modified nucleotide at positions 1-7, 12-27, and 31-36, a GalNAc-conjugated nucleotide at position 28, 29 and 30; and a phosphorothioate linkage between positions 1 and 2;

an antisense strand comprising a 2′-F modified nucleotide at positions 2, 3, 4, 5, 7, 10 and 14, a 2′-OMe at positions 1, 6, 8, 9, 11-13, and 15-22, a phosphorothioate linkage between positions 1 and 2, positions 2 and 3, positions 20 and 21, and positions 21 and 22, and a 5′-terminal nucleotide at position 1 comprising a 4′-phosphate analog, optionally wherein the 5′-terminal nucleotide comprises 5′-methoxyphosphonate-4′-oxy-2′-O-methyluridine [MePhosphonate-4O-mU]; wherein positions 1-20 of the antisense strand form a duplex region with positions 1-20 of the sense strand, wherein positions 21-36 of the sense strand form a stem-loop, wherein positions 27-30 form the loop of the stem-loop, optionally wherein positions 27-30 comprise a tetraloop, wherein positions 21 and 22 of the antisense strand comprise an overhang, and wherein the sense strand and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 886 and 912, respectively;
  • (b) SEQ ID NOs: 887 and 913, respectively;
  • (c) SEQ ID NOs: 910 and 937, respectively;
  • (d) SEQ ID NOs: 888 and 914, respectively;
  • (e) SEQ ID NOs: 889 and 915, respectively;
  • (f) SEQ ID NOs: 890 and 916, respectively;
  • (g) SEQ ID NOs: 891 and 917, respectively;
  • (h) SEQ ID NOs: 877 and 884, respectively;
  • (i) SEQ ID NOs: 878 and 930, respectively;
  • (j) SEQ ID NOs: 876 and 883, respectively;
  • (k) SEQ ID NOs: 875 and 882, respectively;
  • (l) SEQ ID NOs: 892 and 918, respectively;
  • (m) SEQ ID NOs: 893 and 919, respectively;
  • (n) SEQ ID NOs: 894 and 920, respectively;
  • (o) SEQ ID NOs: 904 and 931, respectively;
  • (p) SEQ ID NOs: 895 and 921, respectively;
  • (q) SEQ ID NOs: 905 and 932, respectively;
  • (r) SEQ ID NOs: 896 and 922, respectively;
  • (s) SEQ ID NOs: 911 and 938, respectively;
  • (t) SEQ ID NOs: 906 and 933, respectively;
  • (u) SEQ ID NOs: 897 and 923, respectively;
  • (v) SEQ ID NOs: 907 and 934, respectively;
  • (w) SEQ ID NOs: 908 and 935, respectively;
  • (x) SEQ ID NOs: 903 and 929, respectively;
  • (y) SEQ ID NOs: 901 and 927, respectively;
  • (z) SEQ ID NOs: 874 and 881, respectively;
  • (aa) SEQ ID NOs: 902 and 928, respectively;
  • (bb) SEQ ID NOs: 873 and 880, respectively;
  • (cc) SEQ ID NOs: 872 and 879, respectively;
  • (dd) SEQ ID NOs: 898 and 924, respectively;
  • (ee) SEQ ID NOs: 899 and 925, respectively
  • (ff) SEQ ID NOs: 900 and 926, respectively; and
  • (gg) SEQ ID NOs: 909 and 936, respectively

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 887 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 913. In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 891 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 917. In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 892 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 918. In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 894 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 920. In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 897 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 923. In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 909 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 936.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 948; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 949; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 950; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 951; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 952; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 953; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 948; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 949; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 950; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 951; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 952; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 953; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 948; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 942, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 949; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 943, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 950; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 944, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 951; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 945, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 952; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 946, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 953; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 947, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 948; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 942, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 949; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 943, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 950; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 944, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 951; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 945, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 952; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 946, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is set forth in SEQ ID NO: 953; and (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand and a stem-loop at the 3′terminus, wherein the region of complementarity to the antisense strand is set forth in SEQ ID NO: 947, wherein the stem-loop is set forth as S1-L-S2, wherein S1 is complementary to S2 and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises the modification pattern of:

• • Sense Strand: 5′-mX-S-mX-mX-mX-mX-mX-mX-fX-fX-fX-fX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mX-mX-mX-mX-mX-mX-3′.

Hybridized to:

• • Antisense Strand: 5′-[MePhosphonate-4O-mX]-S-fX-S-fX-S-fX-fX-mX-fX-mX-mX-fX-mX-mX-mX-fX-mX-mX-mX-mX-mX-mX-S-mX-S-mX-3′;wherein mX=2′-O-methyl modified nucleotide, fX=2′-fluoro modified nucleotide, —S—=phosphorothioate linkage, −=phosphodiester linkage, [MePhosphonate-4O-mX]=5′-methoxyphosphonate-4-oxy modified nucleotide, and ademA-GalNAc=GalNAc attached to an adenine nucleotide

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression comprises the modification pattern of

• • Sense Strand: 5′-mX-S-mX-fX-mX-mX-mX-mX-fX-fX-fX-mX-fX-fX-mX-mX-mX-fX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mX-mX-mX-mX-mX-mX-3′.

Hybridized to:

• • Antisense Strand: 5′-[MePhosphonate-4O-mX]-S-fX-S-fX- S-fX-fX-mX-fX-fX-mX-fX-mX-mX-mX-fX-mX-fX-mX-mX-fX-mX-S-mX-S-mX-3′;

wherein mX=2′-O-methyl modified nucleotide, fX=2′-fluoro modified nucleotide, —S—=phosphorothioate linkage, −=phosphodiester linkage, [MePhosphonate-4O-mX]=5′-methoxyphosphonate-4-oxy modified nucleotide, and ademA-GalNAc=GalNAc attached to an adenine nucleotide

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprising a sense strand selected from SEQ ID NOs:774-804 and antisense strand selected from SEQ ID NOs: 819-849. In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 775 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 820. In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 779 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 824. In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 780 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 825. In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 782 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 827. In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 785 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 830. In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprises a sense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 804 and an antisense strand comprising the nucleotide sequence as set forth in SEQ ID NO: 849.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides sense and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 774 and 819, respectively;
  • (b) SEQ ID NOs: 775 and 820, respectively;
  • (c) SEQ ID NOs: 776 and 821, respectively;
  • (d) SEQ ID NOs: 777 and 822, respectively;
  • (e) SEQ ID NOs: 778 and 823, respectively;
  • (f) SEQ ID NOs: 779 and 824, respectively;
  • (g) SEQ ID NOs: 780 and 825, respectively;
  • (h) SEQ ID NOs: 781 and 826, respectively;
  • (i) SEQ ID NOs: 782 and 827, respectively;
  • (j) SEQ ID NOs: 783 and 828, respectively;
  • (k) SEQ ID NOs: 784 and 829, respectively;
  • (l) SEQ ID NOs: 785 and 830, respectively;
  • (m) SEQ ID NOs: 786 and 831, respectively;
  • (n) SEQ ID NOs: 787 and 832, respectively;
  • (o) SEQ ID NOs: 788 and 833, respectively;
  • (p) SEQ ID NOs: 789 and 834, respectively;
  • (q) SEQ ID NOs: 790 and 835, respectively;
  • (r) SEQ ID NOs: 791 and 836, respectively;
  • (s) SEQ ID NOs: 792 and 837, respectively;
  • (t) SEQ ID NOs: 793 and 838, respectively;
  • (u) SEQ ID NOs: 794 and 839, respectively;
  • (v) SEQ ID NOs: 795 and 840, respectively;
  • (w) SEQ ID NOs: 796 and 841, respectively;
  • (x) SEQ ID NOs: 797 and 842, respectively;
  • (y) SEQ ID NOs: 798 and 843, respectively;
  • (z) SEQ ID NOs: 799 and 844, respectively;
  • (aa) SEQ ID NOs: 800 and 845, respectively;
  • (bb) SEQ ID NOs: 801 and 846, respectively;
  • (cc) SEQ ID NOs: 802 and 847, respectively;
  • (dd) SEQ ID NOs: 803 and 848, respectively; and
  • (ee) SEQ ID NOs: 804 and 849, respectively.

In some embodiments, a KHK-targeting dsRNAi oligonucleotide for reducing KHK expression provided by the disclosure comprising a sense strand selected from SEQ ID NOs:805-818 and an antisense strand selected from SEQ ID NOs: 850-863.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides sense and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 805 and 850, respectively;
  • (b) SEQ ID NOs: 806 and 851, respectively;
  • (c) SEQ ID NOs: 807 and 852, respectively;
  • (d) SEQ ID NOs: 808 and 853, respectively;
  • (e) SEQ ID NOs: 809 and 854, respectively;
  • (f) SEQ ID NOs: 810 and 855, respectively;
  • (g) SEQ ID NOs: 811 and 856, respectively;
  • (h) SEQ ID NOs: 812 and 857, respectively;
  • (i) SEQ ID NOs: 813 and 858, respectively;
  • (j) SEQ ID NOs: 814 and 859, respectively;
  • (k) SEQ ID NOs: 815 and 860, respectively;
  • (l) SEQ ID NOs: 816 and 861, respectively;
  • (m) SEQ ID NOs: 817 and 862, respectively and;
  • (n) SEQ ID NOs: 818 and 863, respectively.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides comprise a sense strand comprising SEQ ID NO: 775 and an antisense strand comprising SEQ ID NO: 820, wherein said dsRNA is in the form of a conjugate having the structure as shown in FIG. 10A continuing to FIG. 10B, or pharmaceutically acceptable salts thereof.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides comprise a sense strand comprising SEQ ID NO: 779 and an antisense strand comprising SEQ ID NO: 824, wherein said dsRNA is in the form of a conjugate having as shown in FIG. 11A continuing to FIG. 11B, or pharmaceutically acceptable salts thereof.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides comprise a sense strand comprising SEQ ID NO: 780 and an antisense strand comprising SEQ ID NO: 825, wherein said dsRNA is in the form of a conjugate as depicted in FIG. 12A continuing to FIG. 12B, or pharmaceutically acceptable salts thereof.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides comprise a sense strand comprising SEQ ID NO: 782 and an antisense strand comprising SEQ ID NO: 827, wherein said dsRNA is in the form of a conjugate having the structures depicted in FIG. 13A continuing to FIG. 13B, or pharmaceutically acceptable salts thereof.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides comprise a sense strand comprising SEQ ID NO: 785 and an antisense strand comprising SEQ ID NO: 830, wherein said dsRNA is in the form of a conjugate having the structures depicted in FIG. 14A continuing to FIG. 14B, or pharmaceutically acceptable salts thereof.

In some embodiments, the KHK-targeting dsRNAi oligonucleotides comprise a sense strand comprising SEQ ID NO: 804 and an antisense strand comprising SEQ ID NO: 849, wherein said dsRNA is in the form of a conjugate having the structures depicted in FIG. 15A continuing to FIG. 15B, or pharmaceutically acceptable salts thereof.

Formulations

Various formulations have been developed to facilitate oligonucleotide use. For example, oligonucleotides (e.g., dsRNAi oligonucleotides) can be delivered to a subject or a cellular environment using a formulation that minimizes degradation, facilitates delivery and/or uptake, or provides another beneficial property to the oligonucleotides in the formulation. In some embodiments, provided herein are compositions comprising oligonucleotides (e.g., dsRNAi oligonucleotides) reduce the expression of KHK. Such compositions can be suitably formulated such that when administered to a subject, either into the immediate environment of a target cell or systemically, a sufficient portion of the oligonucleotides enter the cell to reduce KHK expression. Any variety of suitable oligonucleotide formulations can be used to deliver oligonucleotides for the reduction of KHK as disclosed herein. In some embodiments, an oligonucleotide is formulated in buffer solutions such as phosphate buffered saline solutions, liposomes, micellar structures, and capsids. Any of the oligonucleotides described herein may be provided not only as nucleic acids, but also in the form of a pharmaceutically acceptable salt.

Formulations of oligonucleotides with cationic lipids can be used to facilitate transfection of the oligonucleotides into cells. For example, cationic lipids, such as lipofectin, cationic glycerol derivatives, and polycationic molecules (e.g., polylysine), can be used. Suitable lipids include Oligofectamine, Lipofectamine (Life Technologies), NC388 (Ribozyme Pharmaceuticals, Inc., Boulder, Colo.), or FuGene 6 (Roche) all of which can be used according to the manufacturer's instructions.

Accordingly, in some embodiments, a formulation comprises a lipid nanoparticle. In some embodiments, an excipient comprises a liposome, a lipid, a lipid complex, a microsphere, a microparticle, a nanosphere or a nanoparticle, or may be otherwise formulated for administration to the cells, tissues, organs, or body of a subject in need thereof (see, e.g., Remington: THE SCIENCE AND PRACTICE OF PHARMACY, 22nd edition, Pharmaceutical Press, 2013).

In some embodiments, the formulations herein comprise an excipient. In some embodiments, an excipient confers to a composition improved stability, improved absorption, improved solubility and/or therapeutic enhancement of the active ingredient. In some embodiments, an excipient is a buffering agent (e.g., sodium citrate, sodium phosphate, a tris base, or sodium hydroxide) or a vehicle (e.g., a buffered solution, petrolatum, dimethyl sulfoxide, or mineral oil). In some embodiments, an oligonucleotide is lyophilized for extending its shelf-life and then made into a solution before use (e.g., administration to a subject). Accordingly, an excipient in a composition comprising any one of the oligonucleotides described herein may be a lyoprotectant (e.g., mannitol, lactose, polyethylene glycol or polyvinylpyrrolidone) or a collapse temperature modifier (e.g., dextran, Ficoll™ or gelatin).

In some embodiments, a pharmaceutical composition is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral (e.g., intravenous, intramuscular, intraperitoneal, intradermal, subcutaneous), oral (e.g., inhalation), transdermal (e.g., topical), transmucosal and rectal administration.

Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Sterile injectable solutions can be prepared by incorporating the oligonucleotides in a required amount in a selected solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization.

In some embodiments, a composition may contain at least about 0.1% of the therapeutic agent (e.g., a dsRNAi oligonucleotide for reducing KHK expression) or more, although the percentage of the active ingredient(s) may be between about 1% to about 80% or more of the weight or volume of the total composition. Factors such as solubility, bioavailability, biological half-life, route of administration, product shelf life, as well as other pharmacological considerations will be contemplated by one skilled in the art of preparing such pharmaceutical formulations, and as such, a variety of dosages and treatment regimens may be desirable.

Methods of Use

Reducing KHK Expression

In some embodiments, the disclosure provides methods for contacting or delivering to a cell or population of cells an effective amount of oligonucleotides (e.g., dsRNAi oligonucleotides) herein to reduce KHK expression. In some embodiments, a reduction of KHK expression is determined by measuring a reduction in the amount or level of KHK mRNA, KHK protein, or KHK activity in a cell. The methods include those described herein and known to one of ordinary skill in the art.

Methods provided herein are useful in any appropriate cell type. In some embodiments, a cell is any cell that expresses KHK mRNA (e.g., hepatocytes). In some embodiments, the cell is a primary cell obtained from a subject. In some embodiments, the primary cell has undergone a limited number of passages such that the cell substantially maintains its natural phenotypic properties. In some embodiments, a cell to which the oligonucleotide is delivered is ex vivo or in vitro (i.e., can be delivered to a cell in culture or to an organism in which the cell resides).

In some embodiments, the oligonucleotides herein are delivered to a cell or population of cells using a nucleic acid delivery method known in the art including, but not limited to, injection of a solution containing the oligonucleotides, bombardment by particles covered by the oligonucleotides, exposing the cell or population of cells to a solution containing the oligonucleotides, or electroporation of cell membranes in the presence of the oligonucleotides. Other methods known in the art for delivering oligonucleotides to cells may be used, such as lipid-mediated carrier transport, chemical-mediated transport, and cationic liposome transfection such as calcium phosphate, and others.

In some embodiments, reduction of KHK expression is determined by an assay or technique that evaluates one or more molecules, properties, or characteristics of a cell or population of cells associated with KHK expression, or by an assay or technique that evaluates molecules that are directly indicative of KHK expression in a cell or population of cells (e.g., KHK mRNA or KHK protein). In some embodiments, the extent to which an oligonucleotide provided herein reduces KHK expression is evaluated by comparing KHK expression in a cell or population of cells contacted with the oligonucleotide to an appropriate control (e.g., an appropriate cell or population of cells not contacted with the oligonucleotide or contacted with a control oligonucleotide). In some embodiments, a control amount or level of KHK expression in a control cell or population of cells is predetermined, such that the control amount or level need not be measured in every instance the assay or technique is performed. The predetermined level or value can take a variety of forms. In some embodiments, a predetermined level or value can be single cut-off value, such as a median or mean.

In some embodiments, contacting or delivering an oligonucleotide (e.g., dsRNAi oligonucleotides) described herein to a cell or a population of cells results in a reduction in KHK expression in a cell or population of cells not contacted with the oligonucleotide or contacted with a control oligonucleotide. In some embodiments, the reduction in KHK expression is about 1% or lower, about 5% or lower, about 10% or lower, about 15% or lower, about 20% or lower, about 25% or lower, about 30% or lower, about 35% or lower, about 40% or lower, about 45% or lower, about 50% or lower, about 55% or lower, about 60% or lower, about 70% or lower, about 80% or lower, or about 90% or lower relative to a control amount or level of KHK expression. In some embodiments, the control amount or level of KHK expression is an amount or level of KHK mRNA and/or KHK protein in a cell or population of cells that has not been contacted with an oligonucleotide herein. In some embodiments, the effect of delivery of an oligonucleotide to a cell or population of cells according to a method herein is assessed after any finite period or amount of time (e.g., minutes, hours, days, weeks, months). For example, in some embodiments, KHK expression is determined in a cell or population of cells at least about 4 hours, about 8 hours, about 12 hours, about 18 hours, about 24 hours; or at least about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 21 days, about 28 days, about 35 days, about 42 days, about 49 days, about 56 days, about 63 days, about 70 days, about 77 days, or about 84 days or more after contacting or delivering the oligonucleotide to the cell or population of cells. In some embodiments, KHK expression is determined in a cell or population of cells at least about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, or about 6 months or more after contacting or delivering the oligonucleotide to the cell or population of cells.

In some embodiments, an oligonucleotide is delivered in the form of a transgene that is engineered to express in a cell the oligonucleotide or strands comprising the oligonucleotide (e.g., its sense and antisense strands). In some embodiments, an oligonucleotide is delivered using a transgene engineered to express any oligonucleotide disclosed herein. Transgenes may be delivered using viral vectors (e.g., adenovirus, retrovirus, vaccinia virus, poxvirus, adeno-associated virus, or herpes simplex virus) or non-viral vectors (e.g., plasmids or synthetic mRNAs). In some embodiments, transgenes can be injected directly to a subject.

Treatment Methods

The disclosure provides oligonucleotides for use as a medicament, in particular for use in a method for the treatment of diseases, disorders, and conditions associated with expression of KHK. The disclosure also provides oligonucleotides for use, or adaptable for use, to treat a subject (e.g., a human having a disease, disorder or condition associated with KHK expression) that would benefit from reducing KHK expression. In some respects, the disclosure provides oligonucleotides for use, or adapted for use, to treat a subject having a disease, disorder or condition associated with expression of KHK. The disclosure also provides oligonucleotides for use, or adaptable for use, in the manufacture of a medicament or pharmaceutical composition for treating a disease, disorder or condition associated with KHK expression. In some embodiments, the oligonucleotides for use, or adaptable for use, target KHK mRNA and reduce KHK expression (e.g., via the RNAi pathway). In some embodiments, the oligonucleotides for use, or adaptable for use, target KHK mRNA and reduce the amount or level of KHK mRNA, KHK protein and/or KHK activity.

In addition, in some embodiments of the methods herein, a subject having a disease, disorder, or condition associated with KHK expression or is predisposed to the same is selected for treatment with an oligonucleotide (e.g., a double-stranded oligonucleotide) herein. In some embodiments, the method comprises selecting an individual having a marker (e.g., a biomarker) for a disease, disorder, or condition associated with KHK expression or predisposed to the same, such as, but not limited to, KHK mRNA, KHK protein, or a combination thereof. Likewise, and as detailed below, some embodiments of the methods provided by the disclosure include steps such as measuring or obtaining a baseline value for a marker of KHK expression (e.g., KHK), and then comparing such obtained value to one or more other baseline values or values obtained after the subject is administered the oligonucleotide to assess the effectiveness of treatment.

The disclosure also provides methods of treating a subject having, suspected of having, or at risk of developing a disease, disorder or condition associated with a KHK expression with an oligonucleotide provided herein. In some aspects, the disclosure provides methods of treating or attenuating the onset or progression of a disease, disorder or condition associated with KHK expression using the oligonucleotides herein. In other aspects, the disclosure provides methods to achieve one or more therapeutic benefits in a subject having a disease, disorder, or condition associated with KHK expression using the oligonucleotides provided herein. In some embodiments of the methods herein, the subject is treated by administering a therapeutically effective amount of any one or more of the oligonucleotides provided herein. In some embodiments, treatment comprises reducing KHK expression. In some embodiments, the subject is treated therapeutically. In some embodiments, the subject is treated prophylactically.

In some embodiments of the methods herein, one or more oligonucleotides (e.g., dsRNAi oligonucleotides) herein, or a pharmaceutical composition comprising one or more oligonucleotides, is administered to a subject having a disease, disorder or condition associated with KHK expression such that KHK expression is reduced in the subject, thereby treating the subject. In some embodiments, an amount or level of KHK mRNA is reduced in the subject. In some embodiments, an amount or level of KHK protein is reduced in the subject. In some embodiments, an amount or level of KHK activity is reduced in the subject. In some embodiments, an amount or level of triglyceride (TG) (e.g., one or more TG(s) or total TGs) is reduced in the subject. In some embodiments, an amount or level of plasma glucose is reduced in the subject. In some embodiments, an amount or level of blood pressure (e.g., systolic pressure, diastolic pressure, or both) is reduced in the subject. In some embodiments, an amount or level of abdominal fat is reduced in the subject. In some embodiments, an amount or level of cholesterol (e.g., total cholesterol, LDL cholesterol, and/or HDL cholesterol) is reduced in the subject. In some embodiments, an amount or level of liver steatosis is reduced in the subject. In some embodiments, an amount or level of liver fibrosis is reduced in the subject. In some embodiments, the ratio of total cholesterol to HDL cholesterol is altered in the subject. In some embodiments, any combination of the following is reduced or altered in the subject: KHK expression, an amount or level of KHK mRNA, an amount or level of KHK protein, an amount or level of KHK activity, an amount or level of blood glucose, an amount or level of abdominal fat, an amount or level of blood pressure, an amount or level of TG, an amount or level of cholesterol and/or the ratio of total cholesterol to HDL cholesterol, an amount or level of liver steatosis, and amount or level of liver fibrosis.

In some embodiments of the methods herein, an oligonucleotide (e.g., dsRNAi oligonucleotides) herein, or a pharmaceutical composition comprising the oligonucleotide, is administered to a subject having a disease, disorder or condition associated with KHK such that KHK expression is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to KHK expression prior to administration of one or more oligonucleotides or pharmaceutical composition. In some embodiments, KHK expression is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to KHK expression in a subject (e.g., a reference or control subject) not receiving the oligonucleotide or oligonucleotides or pharmaceutical composition or receiving a control oligonucleotide or oligonucleotides, pharmaceutical composition or treatment.

In some embodiments of the methods herein, an oligonucleotide or oligonucleotides herein, or a pharmaceutical composition comprising the oligonucleotide or oligonucleotides, is administered to a subject having a disease, disorder or condition associated with KHK expression such that an amount or level of KHK mRNA is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to the amount or level of KHK mRNA prior to administration of the oligonucleotide or pharmaceutical composition. In some embodiments, an amount or level of KHK mRNA is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to an amount or level of KHK mRNA in a subject (e.g., a reference or control subject) not receiving the oligonucleotide or oligonucleotides or pharmaceutical composition or receiving a control oligonucleotide or oligonucleotides, pharmaceutical composition or treatment.

In some embodiments of the methods herein, an oligonucleotide or oligonucleotides herein, or a pharmaceutical composition comprising the oligonucleotide or oligonucleotides, is administered to a subject having a disease, disorder or condition associated with KHK expression such that an amount or level of KHK protein is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to the amount or level of KHK protein prior to administration of the oligonucleotide or pharmaceutical composition. In some embodiments, an amount or level of KHK protein is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to an amount or level of KHK protein in a subject (e.g., a reference or control subject) not receiving the oligonucleotide or oligonucleotides or pharmaceutical composition or receiving a control oligonucleotide, oligonucleotides or pharmaceutical composition or treatment.

In some embodiments of the methods herein, an oligonucleotide or oligonucleotides (e.g., dsRNAi oligonucleotides) herein, or a pharmaceutical composition comprising the oligonucleotide or oligonucleotides, is administered to a subject having a disease, disorder or condition associated with KHK such that an amount or level of KHK activity/expression is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to the amount or level of KHK activity prior to administration of the oligonucleotide or pharmaceutical composition. In some embodiments, an amount or level of KHK activity is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to an amount or level of KHK activity in a subject (e.g., a reference or control subject) not receiving the oligonucleotide or pharmaceutical composition or receiving a control oligonucleotide, pharmaceutical composition or treatment.

In some embodiments of the methods herein, an oligonucleotide herein, or a pharmaceutical composition comprising the oligonucleotide, is administered to a subject having a disease, disorder or condition associated with KHK expression such that an amount or level of TG (e.g., one or more TGs or total TGs) is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to the amount or level of TG prior to administration of the oligonucleotide or pharmaceutical composition. In some embodiments, an amount or level of TG is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to an amount or level of TG in a subject (e.g., a reference or control subject) not receiving the oligonucleotide or pharmaceutical composition or receiving a control oligonucleotide, pharmaceutical composition or treatment.

Generally, a normal or desirable TG range for a human patient is <150 mg/dL of blood, with <100 being considered ideal. In some embodiments, the patient selected for treatment or treated is identified or determined to have an amount or level of TG of 150 mg/dL. In some embodiments, the patient selected for treatment or treated is identified or determined to have an amount or level of TG in the range of 150 to 199 mg/dL, which is considered borderline high TG levels. In some embodiments, the patient selected for treatment or treated is identified or determined to have an amount or level of TG in the range of 200 to 499 mg/dL, which is considered high TG levels. In some embodiments, the patient selected for treatment or treated is identified or determined to have an amount or level of TG in the range of 500 mg/dL or higher (i.e., 500 mg/dL), which is considered very high TG levels. In some embodiments, the patient selected for treatment or treated is identified or determined to have an amount or level of TG which is 150 mg/dL, 200 mg/dL or 500 mg/dL. In some embodiments, the patient selected for treatment or treated is identified or determined to have an amount of level of TG of 200 to 499 mg/dL, or 500 mg/dL or higher. In some embodiments, the patient selected for treatment or treated is identified or determined to have an amount or level of TG which is 200 mg/dL. In some embodiments of the methods herein, an oligonucleotide (e.g., dsRNAi oligonucleotide) herein, or a pharmaceutical composition comprising the oligonucleotide, is administered to a subject having a disease, disorder or condition associated with KHK expression such that an amount or level of cholesterol (e.g., total cholesterol, LDL cholesterol, and/or HDL cholesterol) is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to the amount or level of cholesterol prior to administration of the oligonucleotide or pharmaceutical composition. In some embodiments, an amount or level of cholesterol is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to an amount or level of cholesterol in a subject (e.g., a reference or control subject) not receiving the oligonucleotide or pharmaceutical composition or receiving a control oligonucleotide, pharmaceutical composition or treatment.

Generally, a normal or desirable cholesterol range (total cholesterol) for an adult human patient is <200 mg/dL of blood. In some embodiments, the patient selected for treatment or treated is identified or determined to have an amount or level of cholesterol of 200 mg/dL. In some embodiments, the patient selected for treatment or treated is identified or determined to have an amount or level of cholesterol in the range of 200 to 239 mg/dL, which is considered borderline high cholesterol levels. In some embodiments, the patient selected for treatment or treated is identified or determined to have an amount or level of cholesterol in the range of 240 mg/dL and higher (i.e., 240 mg/dL), which is considered high cholesterol levels. In some embodiments, the patient selected from treatment or treated is identified or determined to have an amount or level of cholesterol of 200 to 239 mg/dL, or 240 mg/dL or higher. In some embodiments, the patient selected for treatment or treated is identified or determined to have an amount or level of cholesterol which is 200 mg/dL or 240 mg/dL or higher.

In some embodiments of the methods herein, an oligonucleotide herein, or a pharmaceutical composition comprising the oligonucleotide, is administered to a subject having a disease, disorder, or condition associated with KHK expression such that an amount or level of liver fibrosis is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to the amount or level of liver fibrosis prior to administration of the oligonucleotide or pharmaceutical composition. In some embodiments, an amount or level of liver fibrosis is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to an amount or level of liver fibrosis in a subject (e.g., a reference or control subject) not receiving the oligonucleotide or pharmaceutical composition or receiving a control oligonucleotide, pharmaceutical composition or treatment.

In some embodiments of the methods herein, an oligonucleotide herein, or a pharmaceutical composition comprising the oligonucleotide, is administered to a subject having a disease, disorder or condition associated with KHK expression such that an amount or level of liver steatosis is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to the amount or level of liver steatosis prior to administration of the oligonucleotide or pharmaceutical composition. In some embodiments, an amount or level of liver steatosis is reduced in the subject by at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99% when compared to an amount or level of liver steatosis in a subject (e.g., a reference or control subject) not receiving the oligonucleotide or pharmaceutical composition or receiving a control oligonucleotide, pharmaceutical composition or treatment.

Suitable methods for determining KHK expression, the amount or level of KHK mRNA, KHK protein, KHK activity, TG, plasma glucose or cholesterol amount or activity in the subject, or in a sample from the subject, are known in the art. Further, the Examples set forth herein illustrate methods for determining KHK expression.

In some embodiments, KHK expression, the amount or level of KHK mRNA, KHK protein, KHK activity, TG, plasma glucose, or cholesterol, is reduced in a cell (e.g., a hepatocyte), a population or a group of cells (e.g., an organoid), an organ (e.g., liver), blood or a fraction thereof (e.g., plasma), a tissue (e.g., liver tissue), a sample (e.g., a liver biopsy sample), or any other appropriate biological material obtained or isolated from the subject. In some embodiments, KHK expression, the amount or level of KHK mRNA, KHK protein, KHK activity, TG, plasma glucose or cholesterol or any combination thereof, is reduced in more than one type of cell (e.g., a hepatocyte and one or more other type(s) of cell), more than one groups of cells, more than one organ (e.g., liver and one or more other organ(s)), more than one fraction of blood (e.g., plasma and one or more other blood fraction(s)), more than one type of tissue (e.g., liver tissue and one or more other type(s) of tissue), or more than one type of sample (e.g., a liver biopsy sample and one or more other type(s) of biopsy sample).

Generally, a normal or desirable blood sugar level for a human patient is <140 mg/dL. Blood sugar levels between 140 and 199 mg/dL two hours after eating indicates pre-diabetes, and >200 mg/dL indicates diabetes. In some embodiments, the patient selected for treatment or treated is identified or determined to have a level of blood sugar between about 140 mg/dL and about 199 mg/dL, which is considered pre-diabetes. In some embodiments, the patient selected for treatment or treated is identified or determined to have a level of blood sugar 200 mg/dL, which is considered diabetes. In some embodiments of the methods herein, an oligonucleotide (e.g., dsRNAi oligonucleotide) herein, or a pharmaceutical composition comprising the oligonucleotide, is administered to a subject having a disease, disorder or condition associated with KHK expression such that an amount or level of blood sugar is reduced to a normal or pre-diabetes range.

Examples of a disease, disorder or condition associated with KHK expression include, but are not limited to, glucose intolerance, pre-diabetes, type-1 diabetes, type-2 diabetes, metabolic liver diseases, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), drug-induced liver diseases, alcohol-induced liver diseases, infectious agent induced liver diseases, inflammatory liver diseases, immune system dysfunction-mediated liver diseases, dyslipidemia, cardiovascular diseases, restenosis, syndrome X, metabolic syndrome, diabetes, obesity, hypertension, chronic cholangiopathies such as Primary Sclerosing Cholangitis (PSC), Primary Biliary Cholangitis (PBC), biliary atresia, progressive familial intrahepatic cholestasis type 3 (PFIC3), inflammatory bowel diseases, Crohn's disease, ulcerative colitis, liver cancer, hepatocellular carcinoma, gastrointestinal cancer, gastric cancer, colorectal cancer, metabolic disease-induced liver fibrosis or cirrhosis, NAFLD induced fibrosis or cirrhosis, NASH-induced fibrosis or cirrhosis, alcohol-induced liver fibrosis or cirrhosis, drug-induced liver fibrosis or cirrhosis, radiation- or chemotherapy-induced fibrosis or cirrhosis, biliary tract fibrosis, liver fibrosis or cirrhosis due to any chronic cholestatic disease, gut fibrosis of any etiology, Crohn's disease induced fibrosis, ulcerative colitis-induced fibrosis, intestine (e.g. small intestine) fibrosis, colon fibrosis, stomach fibrosis, disease of elevated uric acid (e.g. hyperuricemia, gout), sugar craving, alcohol craving, aldolase B deficiency, hereditary fructose intolerance, chronic kidney disease, diabetic nephropathy, kidney fibrosis, liver failure, liver function loss, coagulopathy, steatohepatitis, disorders of glycemic control, and other KHK-associated metabolic-related disorders and diseases. Of particular interest herein are metabolic syndrome, hypertriglyceridemia, NAFLD, NASH, obesity, or a combination thereof.

Because of their high specificity, the oligonucleotides herein (e.g., dsRNAi oligonucleotides) specifically target mRNAs of target genes of cells and tissue(s), or organs(s) (e.g., liver). In preventing disease, the target gene may be one which is required for initiation or maintenance of the disease or which has been identified as being associated with a higher risk of contracting the disease. In treating disease, the oligonucleotide can be brought into contact with the cells, tissue(s), or organ(s) (e.g., liver) exhibiting or responsible for mediating the disease. For example, an oligonucleotide substantially identical to all or part of a wild-type (i.e., native) or mutated gene associated with a disorder or condition associated with KHK expression may be brought into contact with or introduced into a cell or tissue type of interest such as a hepatocyte or other liver cell.

In some embodiments, the target gene may be a target gene from any mammal, such as a human target. Any gene may be silenced according to the method described herein.

Methods described herein typically involve administering to a subject an effective amount of an oligonucleotide herein (e.g., a dsRNAi oligonucleotide), that is, an amount capable of producing a desirable therapeutic result. A therapeutically acceptable amount may be an amount that can therapeutically treat a disease or disorder. The appropriate dosage for any one subject will depend on certain factors, including the subject's size, body surface area, age, the particular composition to be administered, the active ingredient(s) in the composition, time and route of administration, general health, and other drugs being administered concurrently.

In some embodiments, a subject is administered any one of the compositions herein either enterally (e.g., orally, by gastric feeding tube, by duodenal feeding tube, via gastrostomy or rectally), parenterally (e.g., subcutaneous injection, intravenous injection or infusion, intra-arterial injection or infusion, intraosseous infusion, intramuscular injection, intracerebral injection, intracerebroventricular injection, intrathecal), topically (e.g., epicutaneous, inhalational, via eye drops, or through a mucous membrane), or by direct injection into a target organ (e.g., the liver of a subject). Typically, oligonucleotides herein are administered intravenously or subcutaneously.

As a non-limiting set of examples, the oligonucleotides herein (e.g., dsRNAi oligonucleotides) would typically be administered quarterly (once every three months), bi-monthly (once every two months), monthly or weekly. For example, the oligonucleotides may be administered every week or at intervals of two, or three weeks. Alternatively, the oligonucleotides may be administered daily. In some embodiments, a subject is administered one or more loading doses of the oligonucleotide followed by one or more maintenance doses of the oligonucleotide.

In some embodiments the oligonucleotides herein are administered alone or in combination. In some embodiments the oligonucleotides herein are administered in combination concurrently, sequentially (in any order), or intermittently. For example, two oligonucleotides may be co-administered concurrently. Alternatively, one oligonucleotide may be administered and followed any amount of time later (e.g., one hour, one day, one week or one month) by the administration of a second oligonucleotide.

In some embodiments, the subject to be treated is a human or non-human primate or other mammalian subject. Other exemplary subjects include domesticated animals such as dogs and cats; livestock such as horses, cattle, pigs, sheep, goats, and chickens; and animals such as mice, rats, guinea pigs, and hamsters.

In some embodiments, a single dose of one or more oligonucleotides (e.g., dsRNAi oligonucleotides) herein, or a pharmaceutical composition comprising the oligonucleotide(s), is administered to a subject having a disease, disorder, or condition associated with KHK expression such that an amount or level of KHK mRNA and/or KHK protein, preferably of KHK protein, is reduced in the subject. Said reduction of an amount or level of KHK mRNA and/or KHK protein may be determined by comparison with the amount or level of KHK mRNA and/or KHK protein in a subject (e.g., a reference or control subject) not receiving the oligonucleotide(s) or pharmaceutical composition or receiving one or more control oligonucleotides or pharmaceutical compositions or treatments, or—preferably—by comparison with the amount or level of KHK mRNA and/or KHK protein prior to administration of the oligonucleotide(s) or pharmaceutical composition. Said amount or level of KHK mRNA and/or KHK protein may be determined from liver biopsy samples from the subject. Said single dose may be administered subcutaneously. Said dose of the oligonucleotide(s) may be below 10 mg/kg bodyweight of the subject, e.g. 6 mg/kg or below, in particular from 0.01 mg/kg to 5 mg/kg. Said reduction of an amount or level of KHK mRNA and/or KHK protein may be detectable more than 10 days after the single dose administration of the oligonucleotide(s), e.g. it may remain detectable at day 28, 56, and/or 84 after administration. Said reduction of an amount or level of KHK mRNA and/or KHK protein may be, e.g., at least about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 99% or greater than 99%. In a preferred embodiment, the reduction of an amount or level of KHK mRNA and/or KHK protein remains detectable at day 28, optionally at day 56 and/or 84, after subcutaneous administration of a single dose of one or more oligonucleotides (e.g., dsRNAi oligonucleotides) herein, or a pharmaceutical composition comprising the oligonucleotide(s).

Kits

In some embodiments, the disclosure provides a kit comprising an oligonucleotide herein, and instructions for use. In some embodiments, the kit comprises an oligonucleotide herein, and a package insert containing instructions for use of the kit and/or any component thereof. In some embodiments, the kit comprises, in a suitable container, an oligonucleotide herein, one or more controls, and various buffers, reagents, enzymes and other standard ingredients well known in the art. In some embodiments, the container comprises at least one vial, well, test tube, flask, bottle, syringe, or other container means, into which the oligonucleotide is placed, and in some instances, suitably aliquoted. In some embodiments where an additional component is provided, the kit contains additional containers into which this component is placed. The kits can also include a means for containing the oligonucleotide and any other reagent in close confinement for commercial sale. Such containers may include injection or blow-molded plastic containers into which the desired vials are retained. Containers and/or kits can include labeling with instructions for use and/or warnings.

In some embodiments, a kit comprises an oligonucleotide herein, and a pharmaceutically acceptable carrier, or a pharmaceutical composition comprising the oligonucleotide and instructions for treating or delaying progression of a disease, disorder or condition associated with KHK expression in a subject in need thereof.

Definitions

As used herein, the term “antisense oligonucleotide” encompasses a nucleic acid-based molecule which has a sequence complementary to all or part of the target mRNA, in particular seed sequence thereby capable of forming a duplex with a mRNA. Thus, the term “antisense oligonucleotide”, as used herein, may be referred to as “complementary nucleic acid-based inhibitor”.

As used herein, “approximately” or “about”, as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain embodiments, “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

As used herein, “administer”, “administering”, “administration” and the like refers to providing a substance (e.g., an oligonucleotide) to a subject in a manner that is pharmacologically useful (e.g., to treat a condition in the subject).

As used herein, “attenuate”, “attenuating”, “attenuation” and the like refers to reducing or effectively halting. As a non-limiting example, one or more of the treatments herein may reduce or effectively halt the onset or progression of dyslipidemia/hypertriglyceridemia/hyperlipidemia, NAFLD, NASH, or glucose intolerance in a subject. This attenuation may be exemplified by, for example, a decrease in one or more aspects (e.g., symptoms, tissue characteristics, and cellular, inflammatory or immunological activity, etc.) of dyslipidemia/hypertriglyceridemia/hyperlipidemia, NAFLD, NASH, or glucose intolerance, no detectable progression (worsening) of one or more aspects of dyslipidemia/hypertriglyceridemia/hyperlipidemia, NAFLD, NASH, or glucose intolerance, or no detectable aspects of dyslipidemia/hypertriglyceridemia/hyperlipidemia, NAFLD, NASH, or glucose intolerance in a subject when they might otherwise be expected.

As used herein, “complementary” refers to a structural relationship between two nucleotides (e.g., on two opposing nucleic acids or on opposing regions of a single nucleic acid strand) that permits the two nucleotides to form base pairs with one another. For example, a purine nucleotide of one nucleic acid that is complementary to a pyrimidine nucleotide of an opposing nucleic acid may base pair together by forming hydrogen bonds with one another. In some embodiments, complementary nucleotides can base pair in the Watson-Crick manner or in any other manner that allows for the formation of stable duplexes. In some embodiments, two nucleic acids may have regions of multiple nucleotides that are complementary with each other to form regions of complementarity, as described herein.

As used herein, “deoxyribonucleotide” refers to a nucleotide having a hydrogen in place of a hydroxyl at the 2′ position of its pentose sugar when compared with a ribonucleotide. A modified deoxyribonucleotide is a deoxyribonucleotide having one or more modifications or substitutions of atoms other than at the 2′ position, including modifications or substitutions in or of the sugar, phosphate group or base.

As used herein, “double-stranded oligonucleotide” or “ds oligonucleotide” refers to an oligonucleotide that is substantially in a duplex form. In some embodiments, the complementary base-pairing of duplex region(s) of a double-stranded oligonucleotide is formed between antiparallel sequences of nucleotides of covalently separate nucleic acid strands. In some embodiments, complementary base-pairing of duplex region(s) of a double-stranded oligonucleotide is formed between antiparallel sequences of nucleotides of nucleic acid strands that are covalently linked. In some embodiments, complementary base-pairing of duplex region(s) of a double-stranded oligonucleotide is formed from single nucleic acid strand that is folded (e.g., via a hairpin) to provide complementary antiparallel sequences of nucleotides that base pair together. In some embodiments, a double-stranded oligonucleotide comprises two covalently separate nucleic acid strands that are fully duplexed with one another. However, in some embodiments, a double-stranded oligonucleotide comprises two covalently separate nucleic acid strands that are partially duplexed (e.g., having overhangs at one or both ends). In some embodiments, a double-stranded oligonucleotide comprises antiparallel sequence of nucleotides that are partially complementary, and thus, may have one or more mismatches, which may include internal mismatches or end mismatches.

As used herein, “duplex,” in reference to nucleic acids (e.g., oligonucleotides), refers to a structure formed through complementary base pairing of two antiparallel sequences of nucleotides.

As used herein, “excipient” refers to a non-therapeutic agent that may be included in a composition, for example, to provide or contribute to a desired consistency or stabilizing effect.

As used herein, the phrase “glucose intolerance” refers to a metabolic condition resulting in higher-than-normal levels of blood glucose. Glucose intolerance can include type 1, type 1.5, and type 2 diabetes.

As used herein, “hepatocyte” or “hepatocytes” refers to cells of the parenchymal tissues of the liver. These cells make up about 70%-85% of the liver's mass and manufacture serum albumin, FBN and the prothrombin group of clotting factors (except for Factors 3 and 4). Markers for hepatocyte lineage cells include, but are not limited to, transthyretin (Ttr), glutamine synthetase (Glul), hepatocyte nuclear factor 1a (Hnf1a) and hepatocyte nuclear factor 4a (Hnf4a). Markers for mature hepatocytes may include, but are not limited to, cytochrome P450 (Cyp3a11), fumarylacetoacetate hydrolase (Fah), glucose 6-phosphate (G6p), albumin (Alb) and 002-2F8. See, e.g., Huch et al. (2013) Nature 494:247-50.

As used herein, a “hepatotoxic agent” refers to a chemical compound, virus or other substance that is itself toxic to the liver or can be processed to form a metabolite that is toxic to the liver. Hepatotoxic agents may include, but are not limited to, carbon tetrachloride (0014), acetaminophen (paracetamol), vinyl chloride, arsenic, chloroform, nonsteroidal anti-inflammatory drugs (such as aspirin and phenylbutazone).

As used herein, the term “ketohexokinase” or “KHK” refers to an enzyme, specifically a hepatic fructokinase, that catalyzes the phosphorylation of fructose. The KHK gene encodes two protein isoforms (KHK-A and KHK-C). The two products are generated from the same primary transcript by alternative splicing. The term “KHK” is intended to refer to both isoforms unless stated otherwise. ‘KHK’ may also refer to the gene which encodes the protein.

As used herein, “labile linker” refers to a linker that can be cleaved (e.g., by acidic pH). A “fairly stable linker” refers to a linker that cannot be cleaved.

As used herein, “liver inflammation” or “hepatitis” refers to a physical condition in which the liver becomes swollen, dysfunctional and/or painful, especially as a result of injury or infection, as may be caused by exposure to a hepatotoxic agent. Symptoms may include jaundice (yellowing of the skin or eyes), fatigue, weakness, nausea, vomiting, appetite reduction and weight loss. Liver inflammation, if left untreated, may progress to fibrosis, cirrhosis, liver failure or liver cancer.

As used herein, “liver fibrosis”, “Liver Fibrosis” or “fibrosis of the liver” refers to an excessive accumulation in the liver of extracellular matrix proteins, which could include collagens (I, Ill, and IV), FBN, undulin, elastin, laminin, hyaluronan and proteoglycans resulting from inflammation and liver cell death. Liver fibrosis, if left untreated, may progress to cirrhosis, liver failure or liver cancer.

As used herein, “loop” refers to an unpaired region of a nucleic acid (e.g., oligonucleotide) that is flanked by two antiparallel regions of the nucleic acid that are sufficiently complementary to one another, such that under appropriate hybridization conditions (e.g., in a phosphate buffer, in a cell), the two antiparallel regions, which flank the unpaired region, hybridize to form a duplex (referred to as a “stem”).

As used herein, “Metabolic syndrome’ or “metabolic liver disease” refers to a disorder characterized by a cluster of associated medical conditions and associated pathologies including, but not limited to the following medical conditions: abdominal obesity, elevated blood pressure, elevated fasting plasma glucose, high serum triglycerides, liver fibrosis, and low levels of high-density lipoprotein (HDL) levels. As used herein, the term metabolic syndrome or metabolic liver disease may encompass a wide array of direct and indirect manifestations, diseases and pathologies associated with metabolic syndrome and metabolic liver disease, with an expanded list of conditions used throughout the document.

As used herein, “modified internucleotide linkage” refers to an internucleotide linkage having one or more chemical modifications when compared with a reference internucleotide linkage comprising a phosphodiester bond. In some embodiments, a modified nucleotide is a non-naturally occurring linkage. Typically, a modified internucleotide linkage confers one or more desirable properties to a nucleic acid in which the modified internucleotide linkage is present. For example, a modified internucleotide linkage may improve thermal stability, resistance to degradation, nuclease resistance, solubility, bioavailability, bioactivity, reduced immunogenicity, etc.

As used herein, “modified nucleotide” refers to a nucleotide having one or more chemical modifications when compared with a corresponding reference nucleotide selected from: adenine ribonucleotide, guanine ribonucleotide, cytosine ribonucleotide, uracil ribonucleotide, adenine deoxyribonucleotide, guanine deoxyribonucleotide, cytosine deoxyribonucleotide and thymidine deoxyribonucleotide. In some embodiments, a modified nucleotide is a non-naturally occurring nucleotide. In some embodiments, a modified nucleotide has one or more chemical modification in its sugar, nucleobase and/or phosphate group. In some embodiments, a modified nucleotide has one or more chemical moieties conjugated to a corresponding reference nucleotide. Typically, a modified nucleotide confers one or more desirable properties to a nucleic acid in which the modified nucleotide is present. For example, a modified nucleotide may improve thermal stability, resistance to degradation, nuclease resistance, solubility, bioavailability, bioactivity, reduced immunogenicity, etc.

As used herein, “nicked tetraloop structure” refers to a structure of a RNAi oligonucleotide that is characterized by separate sense (passenger) and antisense (guide) strands, in which the sense strand has a region of complementarity with the antisense strand, and in which at least one of the strands, generally the sense strand, has a tetraloop configured to stabilize an adjacent stem region formed within the at least one strand.

As used herein, “oligonucleotide” refers to a short nucleic acid (e.g., less than about 100 nucleotides in length). An oligonucleotide may be single-stranded (ss) or ds. An oligonucleotide may or may not have duplex regions. As a set of non-limiting examples, an oligonucleotide may be, but is not limited to, a small interfering RNA (siRNA), microRNA (miRNA), short hairpin RNA (shRNA), dicer substrate interfering RNA (DsiRNA), antisense oligonucleotide, short siRNA or ss siRNA. In some embodiments, a double-stranded (dsRNA) is an RNAi oligonucleotide.

As used herein, “overhang” (or “overhang sequence”) refers to terminal non-base pairing nucleotide(s) resulting from one strand or region extending beyond the terminus of a complementary strand with which the one strand or region forms a duplex. In some embodiments, an overhang comprises one or more unpaired nucleotides extending from a duplex region at the 5′ terminus or 3′ terminus of a dsRNA. In certain embodiments, the overhang is a 3′ or 5′ overhang on the antisense strand or sense strand of a dsRNA.

As used herein, “phosphate analog” refers to a chemical moiety that mimics the electrostatic and/or steric properties of a phosphate group. In some embodiments, a phosphate analog is positioned at the 5′ terminal nucleotide of an oligonucleotide in place of a 5′-phosphate, which is often susceptible to enzymatic removal. In some embodiments, a 5′ phosphate analog contains a phosphatase-resistant linkage. Examples of phosphate analogs include, but are not limited to, 5′ phosphonates, such as 5′ methylene phosphonate (5′-MP) and 5′-(E)-vinylphosphonate (5′-VP). In some embodiments, an oligonucleotide has a phosphate analog at a 4′-carbon position of the sugar (referred to as a “4′-phosphate analog”) at a 5′-terminal nucleotide. An example of a 4′-phosphate analog is oxymethylphosphonate, in which the oxygen atom of the oxymethyl group is bound to the sugar moiety (e.g., at its 4′-carbon) or analog thereof. See, e.g., US Patent Publication No. 2019-0177729. Other modifications have been developed for the 5′ end of oligonucleotides (see, e.g., Intl. Patent Application No. WO 2011/133871; U.S. Pat. No. 8,927,513; and Prakash et al. (2015) NUCLEIC ACIDS RES. 43:2993-3011).

As used herein, “reduced expression” of a gene (e.g., KHK) refers to a decrease in the amount or level of RNA transcript (e.g., KHK mRNA) or protein encoded by the gene and/or a decrease in the amount or level of activity of the gene in a cell, a population of cells, a sample, or a subject, when compared to an appropriate reference (e.g., a reference cell, population of cells, sample or subject). For example, the act of contacting a cell with an oligonucleotide herein (e.g., an oligonucleotide comprising an antisense strand having a nucleotide sequence that is complementary to a nucleotide sequence comprising KHK mRNA) may result in a decrease in the amount or level of KHK mRNA, protein and/or activity (e.g., via degradation of KHK mRNA by the RNAi pathway) when compared to a cell that is not treated with the dsRNA. Similarly, and as used herein, “reducing expression” refers to an act that results in reduced expression of a gene (e.g., KHK).

As used herein, “reduction of KHK expression” refers to a decrease in the amount or level of KHK mRNA, KHK protein and/or KHK activity in a cell, a population of cells, a sample or a subject when compared to an appropriate reference (e.g., a reference cell, population of cells, sample, or subject).

As used herein, “region of complementarity” refers to a sequence of nucleotides of a nucleic acid (e.g., a dsRNA) that is sufficiently complementary to an antiparallel sequence of nucleotides to permit hybridization between the two sequences of nucleotides under appropriate hybridization conditions (e.g., in a phosphate buffer, in a cell, etc.). In some embodiments, an oligonucleotide herein comprises a targeting sequence having a region of complementarity to a mRNA target sequence. In some embodiments, the region of complementarity is full complementary. In some embodiments, the region of complementarity is partially complementary (e.g., up to 3 nucleotide mismatches).

As used herein, “ribonucleotide” refers to a nucleotide having a ribose as its pentose sugar, which contains a hydroxyl group at its 2′ position. A modified ribonucleotide is a ribonucleotide having one or more modifications or substitutions of atoms other than at the 2′ position, including modifications or substitutions in or of the ribose, phosphate group or base.

As used herein, “RNAi oligonucleotide” refers to either (a) a double-stranded oligonucleotide having a sense strand (passenger) and antisense strand (guide), in which the antisense strand or part of the antisense strand is used by the Argonaute 2 (Ago2) endonuclease in the cleavage of a target mRNA (e.g., KHK mRNA) or (b) a single-stranded oligonucleotide having a single antisense strand, where that antisense strand (or part of that antisense strand) is used by the Ago2 endonuclease in the cleavage of a target mRNA (e.g., KHK mRNA).

As used herein, “strand” refers to a single, contiguous sequence of nucleotides linked together through internucleotide linkages (e.g., phosphodiester linkages or phosphorothioate linkages). In some embodiments, a strand has two free ends (e.g., a 5′ end and a 3′ end).

As used herein, “subject” means any mammal, including mice, rabbits, and humans. In one embodiment, the subject is a human or NHP. Moreover, “individual” or “patient” may be used interchangeably with “subject.”

As used herein, “synthetic” refers to a nucleic acid or other molecule that is artificially synthesized (e.g., using a machine (e.g., a solid-state nucleic acid synthesizer)) or that is otherwise not derived from a natural source (e.g., a cell or organism) that normally produces the molecule.

As used herein, “targeting ligand” refers to a molecule (e.g., a carbohydrate, amino sugar, cholesterol, polypeptide, or lipid) that selectively binds to a cognate molecule (e.g., a receptor) of a tissue or cell of interest and that is conjugatable to another substance for purposes of targeting the other substance to the tissue or cell of interest. For example, in some embodiments, a targeting ligand may be conjugated to an oligonucleotide for purposes of targeting the oligonucleotide to a specific tissue or cell of interest. In some embodiments, a targeting ligand selectively binds to a cell surface receptor. Accordingly, in some embodiments, a targeting ligand when conjugated to an oligonucleotide facilitates delivery of the oligonucleotide into a particular cell through selective binding to a receptor expressed on the surface of the cell and endosomal internalization by the cell of the complex comprising the oligonucleotide, targeting ligand and receptor. In some embodiments, a targeting ligand is conjugated to an oligonucleotide via a linker that is cleaved following or during cellular internalization such that the oligonucleotide is released from the targeting ligand in the cell.

As used herein, “tetraloop” refers to a loop that increases stability of an adjacent duplex formed by hybridization of flanking sequences of nucleotides. The increase in stability is detectable as an increase in melting temperature (T_m) of an adjacent stem duplex that is higher than the T_m of the adjacent stem duplex expected, on average, from a set of loops of comparable length consisting of randomly selected sequences of nucleotides. For example, a tetraloop can confer a T_m of at least about 50° C., at least about 55° C., at least about 56° C., at least about 58° C., at least about 60° C., at least about 65° C. or at least about 75° C. in 10 mM Na₂HPO₄ to a hairpin comprising a duplex of at least 2 base pairs (bp) in length. In some embodiments, a tetraloop can confer a Tm of at least about 50° C., at least about 55° C., at least about 56° C., at least about 58° C., at least about 60° C., at least about 65° C. or at least about 75° C. in 10 mM NaH₂PO₄ to a hairpin comprising a duplex of at least 2 base pairs (bp) in length. In some embodiments, a tetraloop may stabilize a bp in an adjacent stem duplex by stacking interactions. In addition, interactions among the nucleotides in a tetraloop include, but are not limited to, non-Watson-Crick base pairing, stacking interactions, hydrogen bonding and contact interactions (Cheong et al. (1990) NATURE 346:680-82; Heus & Pardi (1991) SCIENCE 253:191-94). In some embodiments, a tetraloop comprises or consists of 3 to 6 nucleotides and is typically 4 to 5 nucleotides. In certain embodiments, a tetraloop comprises or consists of 3, 4, 5 or 6 nucleotides, which may or may not be modified (e.g., which may or may not be conjugated to a targeting moiety). In one embodiment, a tetraloop consists of 4 nucleotides. Any nucleotide may be used in the tetraloop and standard IUPAC-IUB symbols for such nucleotides may be used as described in Cornish-Bowden (1985) Nucleic Acids Res. 13:3021-3030. For example, the letter “N” may be used to mean that any base may be in that position, the letter “R” may be used to show that A (adenine) or G (guanine) may be in that position, and “B” may be used to show that C (cytosine), G (guanine), or T (thymine) may be in that position. Examples of tetraloops include the UNCG family of tetraloops (e.g., UUCG), the GNRA family of tetraloops (e.g., GAAA), and the CUUG tetraloop (Woese et al. (1990) PROC. NATL. ACAD. SCI. USA 87:8467-8471; Antao et al. (1991) NUCLEIC ACIDS RES. 19:5901-5905). Examples of DNA tetraloops include the d(GNNA) family of tetraloops (e.g., d(GTTA), the d(GNRA)) family of tetraloops, the d(GNAB) family of tetraloops, the d(CNNG) family of tetraloops, and the d(TNCG) family of tetraloops (e.g., d(TTCG)). See, e.g., Nakano et al. (2002) BIOCHEM. 41:14281-92; Okabe et al. (2000) NIPPON KAGAKKAI KOEN YOKOSHU 78:731. In some embodiments, the tetraloop is contained within a nicked tetraloop structure.

As used herein, “treat” or “treating” refers to the act of providing care to a subject in need thereof, for example, by administering a therapeutic agent (e.g., an oligonucleotide herein) to the subject, for purposes of improving the health and/or well-being of the subject with respect to an existing condition (e.g., a disease, disorder) or to prevent or decrease the likelihood of the occurrence of a condition. In some embodiments, treatment involves reducing the frequency or severity of at least one sign, symptom or contributing factor of a condition (e.g., disease, disorder) experienced by a subject.

EXAMPLES

While the disclosure has been described with reference to the specific embodiments set forth in the following Examples, it should be understood by those skilled in the art that various changes may be made, and equivalents may be substituted without departing from the true spirit and scope of the disclosure. Further, the following Examples are offered by way of illustration and are not intended to limit the scope of the disclosure in any manner. In addition, modifications may be made to adapt to a situation, material, composition of matter, process, process step or steps, to the objective, spirit, and scope of the disclosure. All such modifications are intended to be within the scope of the disclosure. Standard techniques well known in the art or the techniques specifically described below were utilized.

RNAi agents targeting KHK have been described and tested in vitro (e.g., WO 2015123264 and WO 2020060986). The following studies describe the identification of novel dsRNAi agents useful for reducing or inhibiting KHK expression based on in vitro and in vivo screening, including studies in non-human primates. The novel dsRNAi agents comprise 36mer sense strands and 22mer antisense strands with a stem loop having a nicked tetraloop conjugated to GalNAc moieties at the 3′end of the sense strand for reducing KHK mRNA. The presence of a nick within the stem loop provides a precut antisense strand to form a pre-processed binding substrate for the Dicer enzyme, allowing Dicer to efficiently bind and hand off the double stranded molecule to Ago2. The tetraloop provides a thermodynamically stabilizing element to prevent the loop from opening and exposing the 5′-end of the antisense strand and the 3′-end of the sense strand, thereby providing increased nuclease resistance. Accordingly, the present dsRNAi agents are particularly useful for inhibiting KHK expression in vitro and in vivo as described in the following examples.

In comparison to dsRNAi agents described in the prior art, the dsRNAi agents presented herein may, in particular, show improved in vitro and/or in vivo reduction or inhibition of KHK expression as determined on the KHK mRNA and/or KHK protein level. Such improvement may relate to the size and/or duration of the inhibitory action. Thus, for instance, for medical uses of the dsRNAi agents according to this invention, lower doses and/or lower dose frequencies may be applicable. Also, dsRNAi agents presented herein may benefit from advantageous safety and tolerability features like high specificity, low off-target effects or reduced immunogenicity.

Example 1: Preparation of Double-Stranded RNAi Oligonucleotides

Oligonucleotide Synthesis and Purification

The double-stranded RNAi (dsRNA) oligonucleotides described in the foregoing Examples are chemically synthesized using methods described herein. Generally, dsRNAi oligonucleotides are synthesized using solid phase oligonucleotide synthesis methods as described for 19-23mer siRNAs (see, e.g., Scaringe et al. (1990) NUCLEIC ACIDS RES. 18:5433-5441 and Usman et al. (1987) J. AM. CHEM. SOC. 109:7845-7845; see also, U.S. Pat. Nos. 5,804,683; 5,831,071; 5,998,203; 6,008,400; 6,111,086; 6,117,657; 6,353,098; 6,362,323; 6,437,117 and 6,469,158) in addition to using known phosphoramidite synthesis (see, e.g. Hughes and Ellington (2017) COLD SPRING HARB PERSPECT BIOL. 9(1):a023812; Beaucage S. L., Caruthers M. H., Studies on Nucleotide Chemistry V: Deoxynucleoside Phosphoramidites—A New Class of Key Intermediates for Deoxypolynucleotide Synthesis, TETRAHEDRON LETT. 1981; 22:1859-62. doi: 10.1016/S0040-4039(01)90461-7). dsRNAi oligonucleotides having a 19mer core sequence were formatted into constructs having a 25mer sense strand and a 27mer antisense strand to allow for processing by the RNAi machinery. The 19mer core sequence is complementary to a region in the KHK mRNA.

Individual RNA strands were synthesized and HPLC purified according to standard methods (Integrated DNA Technologies; Coralville, Iowa). For example, RNA oligonucleotides were synthesized using solid phase phosphoramidite chemistry, deprotected and desalted on NAP-5 columns (Amersham Pharmacia Biotech; Piscataway, N.J.) using standard techniques (Damha & Olgivie (1993) METHODS MOL. BIOL. 20:81-114; Wincott et al. (1995) NUCLEIC ACIDS RES. 23:2677-84). The oligomers were purified using ion-exchange high performance liquid chromatography (IE-HPLC) on an Amersham Source 15Q column (1.0 cm×25 cm; Amersham Pharmacia Biotech) using a 15 min step-linear gradient. The gradient varied from 90:10 Buffers A:B to 52:48 Buffers A:B, where Buffer A is 100 mM Tris pH 8.5 and Buffer B is 100 mM Tris pH 8.5, 1 M NaCl. Samples were monitored at 260 nm and peaks corresponding to the full-length oligonucleotide species were collected, pooled, desalted on NAP-5 columns, and lyophilized.

The purity of each oligomer was determined by capillary electrophoresis (CE) on a Beckman PACE 5000 (Beckman Coulter, Inc.; Fullerton, Calif.). The CE capillaries have a 100 μm inner diameter and contain ssDNA 100R Gel (Beckman-Coulter). Typically, about 0.6 nmole of oligonucleotide was injected into a capillary, run in an electric field of 444 V/cm, and was detected by UV absorbance at 260 nm. Denaturing Tris-Borate-7 M-urea running buffer was purchased from Beckman-Coulter. Oligoribonucleotides were obtained that were at least 90% pure as assessed by CE for use in experiments described below. Compound identity was verified by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectroscopy on a Voyager DE™ Biospectometry Work Station (Applied Biosystems; Foster City, Calif.) following the manufacturer's recommended protocol. Relative molecular masses of all oligomers were obtained, often within 0.2% of expected molecular mass.

Preparation of Duplexes

Single strand RNA oligomers were resuspended (e.g., at 100 μM concentration) in duplex buffer consisting of 100 mM potassium acetate, 30 mM HEPES, pH 7.5. Complementary sense and antisense strands were mixed in equal molar amounts to yield a final solution of, for example, 50 μM duplex. Samples were heated to 100° C. for 5′ in RNA buffer (IDT) and were allowed to cool to room temperature before use. The dsRNA oligonucleotides were stored at −20° C. Single strand RNA oligomers were stored lyophilized or in nuclease-free water at −80° C.

Example 2: Generation of KHK-Targeting Double-Stranded (DS) RNAi Oligonucleotides

Identification of KHK mRNA Target Sequences

Ketohexokinase (KHK) is an enzyme involved in fructose metabolism. KHK has two isoforms, differing by one alternative exon, with distinct substrates and mechanisms of action. The isoform KHK-A is encoded by Exon 3A whereas the KHK-C isoform is encoded by Exon 3C. To generate RNAi oligonucleotide inhibitors of KHK-A and KHK-C expression, a computer-based algorithm was used to computationally identify KHK mRNA target sequences suitable for assaying inhibition of KHK expression by the RNAi pathway. The algorithm provided RNAi oligonucleotide guide (antisense) strand sequences each having a region of complementarity to a suitable KHK target sequence of human KHK mRNA (e.g., SEQ ID NO: 1; Table 1). Some of the guide strand sequences identified by the algorithm were also complementary to the corresponding KHK target sequence of monkey and/or mouse KHK mRNA (SEQ ID NO: 2 and 3, respectively; Table 1). KHK RNAi oligonucleotides comprising a region of complementarity to homologous KHK mRNA target sequences with nucleotide sequence similarity are predicted to have the ability to target homologous KHK mRNAs.

TABLE 1
Sequences of Human, Monkey and Mouse KHK mRNA
Species	Ref Seq #	SEQ ID NO
Human (Hs)	NM_006488.3	1
Cynomolgus monkey (Mf)	XM_005576322.2	2
Mouse (Mm)	NM_008439.4	3

RNAi oligonucleotides (formatted as DsiRNA oligonucleotides) were generated as described in Example 1 for evaluation in vitro. Each DsiRNA was generated with the same modification pattern, and each with a unique guide strand having a region of complementarity to a KHK target sequence identified by the algorithm (Table 2). Modifications for the sense and anti-sense DsiRNA included the following (X− any nucleotide; m-2′-O-methyl modified nucleotide; r-ribosyl modified nucleotide):

Sense Strand:
rXmXrXmXrXrXrXrXrXrXrXrXrXmXrXmXrXrXrXrXrXrXrXXX
Anti-sense Strand:
mXmXmXmXrXrXrXrXrXrXmXrXmXrXrXrXrXrXrXrXrXrXmXrX
mXmXmX

In Vitro Cell-Based Assays

The ability of each of the modified DsiRNA in Table 2 to reduce KHK mRNA was measured using in vitro cell-based assays. Briefly, human hepatoma (Hep3B) cells expressing endogenous human KHK gene were transfected with each of the DsiRNAs listed in Table 2 (Sense Strand SEQ ID NOs: 4-387) at 1 nM in separate wells of a multi-well cell-culture plate. Cells were maintained for 24 hours following transfection with the modified DsiRNA, and then the amount of remaining KHK mRNA from the transfected cells was determined using TAQMAN®-based qPCR assays. Two qPCR assays, a 3′ assay (Forward-1026; TGGAGGTGGAGAAGCCA, Reverse-1157; GACCATACAAGCCCCTCAAG, Probe-1080; TGGTGTTTGTCAGCAAAGATGTGGC) and a 5′ assay (Forward-496; AGGAAGCTCTGGGAGTA, Reverse-596; CCTCCTTAGGGTACTTGTC, Probe-518; ATGGAAGAGAAGCAGATCCTGTGCG) were used to determine KHK mRNA levels as measured using PCR probes conjugated to 6-carboxy-fluorescein (FAM). Each primer pair (KHK-825 for KHK-C isoform, NM_006488.3) and KHK-All (both isoforms) (KHK-F495, KHK-F1026 for KHK-All (both isoforms) was assayed for % remaining RNA as shown in Table 2 and FIG. 1. DsiRNAs resulting in less than or equal to 10% KHK mRNA remaining in DsiRNA-transfected cells when compared to mock-transfected cells were considered DsiRNA “hits”. The Hep3B cell-based assay evaluating the ability of the DsiRNAs listed in Table 2 to inhibit KHK expression identified several candidate DsiRNAs.

Taken together, these results show that DsiRNAs designed to target human KHK mRNA inhibit KHK expression in cells, as determined by a reduced amount of KHK mRNA in DsiRNA-transfected cells relative to control cells. These results demonstrate that the nucleotide sequences comprising the DsiRNA are useful for generating RNAi oligonucleotides to inhibit KHK expression. Further, these results demonstrate that multiple KHK mRNA target sequences are suitable for the RNAi-mediated inhibition of KHK expression.

TABLE 2
Analysis of KHK mRNA in HepB3 cells
SEQ ID NO	SEQ ID NO		KHK-F495	KHK-F1026	KHK-825	% average
(Sense	(Anti-sense		%		%		%		of all
Strand)	Strand)	DsiRNA name	remaining	SEM	remaining	SEM	remaining	SEM	assays
4	388	KHK-115-154	28.3	2.4	33.1	2.7	30.22	6.148	30.5
5	389	KHK-116-155	27.8	5.9	33.3	6.5	25.691	8.728	28.9
6	390	KHK-117-156	26.7	9.9	47.5	14.6	32.656	13.903	35.6
7	391	KHK-118-157	43.7	6.3	63.9	6.4	47.299	13.144	51.6
8	392	KHK-119-158	34.6	8.6	57.5	14.7	22.791	11.895	38.3
9	393	KHK-120-159	31.4	3.8	49.7	3.5	40.388	5.289	40.5
10	394	KHK-121-160	35.8	4.4	53.2	5.7	43.359	8.043	44.1
11	395	KHK-122-161	41.8	3.7	67.6	6.5	39.75	5.587	49.7
12	396	KHK-123-162	49.5	3.7	66.7	4.6	41.386	7.102	52.5
13	397	KHK-124-163	30.9	2.5	49.6	4.4	40.966	7.589	40.5
14	398	KHK-125-164	39.5	3.7	52.5	5.1	43.748	9.465	45.3
15	399	KHK-126-165	37.1	2.8	57.9	5.2	53.25	7.484	49.4
16	400	KHK-127-166	38.6	3.1	63.0	4.3	45.393	5.073	49.0
17	401	KHK-128-167	31.1	2.6	44.6	3.3	47.943	5.962	41.2
18	402	KHK-179	28.9	23.1	81.2	21.2	62.845	5.928	57.6
19	403	KHK-181-220	43.4	5.3	57.6	5.6	56.679	10.519	52.5
20	404	KHK-182-221	33.8	1.8	40.4	3.4	50.153	10.698	41.5
21	405	KHK-183-222	48.9	3.5	57.7	5.4	58.074	7.904	54.9
22	406	KHK-184-223	37.0	2.6	38.5	3.0	49.417	4.396	41.6
23	407	KHK-185-224	34.3	4.0	44.8	4.3	53.186	7.765	44.1
24	408	KHK-186-225	36.1	4.9	42.4	5.0	51.087	6.631	43.2
25	409	KHK-187-226	48.5	3.9	48.2	4.5	81.75	6.116	59.5
26	410	KHK-188-227	50.9	2.8	50.5	5.6	75.858	6.936	59.1
27	411	KHK-431-470	38.7	2.5	42.1	3.8	52.142	5.549	44.3
28	412	KHK-432-471	74.1	38.8	65.5	11.0	50.476	11.385	63.3
29	413	KHK-433-472	53.1	8.6	72.4	9.0	47.428	12.036	57.7
30	414	KHK-507-545-376-218	23.8	3.1	31.1	2.5	84.241	10.972	46.4
31	415	KHK-508-546-377-219	15.7	1.8	43.7	2.6	52.076	6.767	37.2
32	416	KHK-509-547-378-220	10.9	2.4	24.5	4.2	25.526	5.12	20.3
33	417	KHK-510-548-379-221	9.1	3.2	15.0	5.0	15.853	4.385	13.3
34	418	KHK-511-549-380-222	10.4	3.0	23.8	3.5	17.016	6.998	17.1
35	419	KHK-512-550-381-223	6.5	2.2	28.8	8.3	33.059	5.266	22.8
36	420	KHK-513-551-382-224	25.4	8.6	25.2	3.2	22.514	5.225	24.4
37	421	KHK-514-552-383-225	16.4	4.8	33.5	11.4	45.353	14.944	31.7
38	422	KHK-515-553-384-226	66.1	4.5	47.0	3.6	150.28	13.471	87.8
39	423	KHK-516-554-385-227	10.6	2.6	19.4	5.6	27.145	7.112	19.0
40	424	KHK-517-555-386-228	12.0	3.4	24.7	6.3	24.382	9.127	20.4
41	425	KHK-518-556-387-229	15.9	5.6	23.1	6.5	16.128	6.295	18.4
42	426	KHK-520-558-389-231	8.9	1.9	15.6	3.4	20.589	4.844	15.0
43	427	KHK-521-559-390-232	5.0	2.9	23.6	6.4	23.218	8.661	17.3
44	428	KHK-522-560-391-233	9.7	1.7	12.3	2.7	12.226	3.342	11.4
45	429	KHK-541-579	17.8	1.5	26.0	2.6	27.272	5.808	23.7
46	430	KHK-544-582	22.9	2.7	18.4	1.9	59.795	8.985	33.7
47	431	KHK-546-584	30.6	5.8	42.2	7.1	35.433	9.834	36.1
48	432	KHK-547-585	12.2	1.9	17.8	2.6	20.431	7.549	16.8
49	433	KHK-548-586	11.8	2.3	25.1	7.8	7.622	2.199	14.8
50	434	KHK-549-587	12.2	2.0	15.0	3.6	14.783	3.155	14.0
51	435	KHK-550-588	15.9	6.2	23.7	8.1	18.365	7.143	19.3
52	436	KHK-551-589	24.7	4.2	34.8	6.1	14.11	5.994	24.5
53	437	KHK-552-590	24.6	4.3	30.5	5.3	22.483	8.443	25.9
54	438	KHK-553-591	16.1	1.6	27.3	4.3	21.753	4.168	21.7
55	439	KHK-554-592	18.9	6.7	26.3	9.0	20.976	8.622	22.1
56	440	KHK-555-593	22.2	7.1	27.2	10.4	24.612	15.246	24.7
57	441	KHK-556-594	25.9	7.5	32.3	9.9	24.205	11.232	27.5
58	442	KHK-557-595	15.1	1.9	18.9	2.2	29.27	5.8	21.1
59	443	KHK-558-596	25.6	8.0	24.2	11.2	29.227	13.255	26.4
60	444	KHK-559-597	21.3	1.9	29.7	1.8	23.268	4.973	24.8
61	445	KHK-560-598	58.6	2.8	77.3	4.2	61.287	4.768	65.7
62	446	KHK-561-599	14.7	2.2	28.9	3.3	23.131	4.888	22.3
63	447	KHK-562-600	15.1	5.1	21.2	6.5	22.347	6.503	19.6
64	448	KHK-563-601	34.7	1.7	47.5	2.5	63.417	8.195	48.5
65	449	KHK-564-602	19.2	1.8	30.1	2.4	44.737	9.177	31.3
66	450	KHK-565-603	55.7	3.1	73.0	5.6	106.349	11.887	78.3
67	451	KHK-566-604	19.7	1.0	30.1	2.8	31.606	2.125	27.1
68	452	KHK-567-605	9.9	0.5	15.4	1.3	14.15	2.218	13.2
69	453	KHK-568-606	15.6	1.0	19.6	1.1	21.246	2.662	18.8
70	454	KHK-569-607	45.3	5.4	53.3	5.9	42.462	12.958	47.0
71	455	KHK-570-608	14.6	1.4	21.6	2.3	24.089	3.615	20.1
72	456	KHK-571-609	15.4	1.8	17.4	1.9	28.812	4.966	20.6
73	457	KHK-572-610	27.2	1.2	33.3	2.2	38.47	4.931	33.0
74	458	KHK-573-611	20.6	1.7	23.1	2.1	50.396	5.757	31.4
75	459	KHK-574-612	19.8	1.6	27.3	2.2	29.683	5.49	25.6
76	460	KHK-575-613	31.1	1.9	31.2	2.4	37.359	5.282	33.2
77	461	KHK-576-614	30.0	4.0	32.2	1.9	19.014	3.763	27.1
78	462	KHK-577-615	20.8	1.9	31.6	2.8	22.807	5.999	25.1
79	463	KHK-638-676	25.1	2.0	27.0	2.0	38.226	3.255	30.1
80	464	KHK-641-679	29.0	1.7	33.5	2.0	30.662	5.567	31.0
81	465	KHK-642-680	22.6	2.6	31.3	2.7	12.983	3.632	22.3
82	466	KHK-643-681	39.1	2.1	41.5	2.4	38.644	6.762	39.7
83	467	KHK-644-682	25.3	3.2	34.7	7.5	11.343	4.407	23.8
84	468	KHK-645-683	15.9	5.0	21.0	3.1	21.433	1.586	19.5
85	469	KHK-646-684	22.8	1.8	30.8	2.2	13.876	3.967	22.5
86	470	KHK-647-685	38.8	1.5	41.9	2.4	32.316	6.062	37.7
87	471	KHK-650-688	40.6	2.1	42.3	3.2	41.675	12.029	41.5
88	472	KHK-676-714	56.0	2.4	52.8	2.8	71.792	10.911	60.2
89	473	KHK-713-722	71.9	3.5	78.9	3.4	49.345	10.259	66.7
90	474	KHK-826-835	87.0	6.4	101.5	5.1	76.866	14.977	88.5
91	475	KHK-827-836	62.3	2.7	98.9	8.8	29.087	3.652	63.4
92	476	KHK-829-838	100.3	4.5	86.8	5.3	17.67	3.397	68.3
93	477	KHK-830-839	67.4	3.2	81.8	3.6	17.686	2.276	55.6
94	478	KHK-831-840	61.7	3.4	69.8	5.2	13.313	3.649	48.3
95	479	KHK-832-841	82.3	4.4	80.0	4.4	34.692	4.011	65.7
96	480	KHK-857-895	73.2	10.7	53.0	13.9	88.908	21.326	71.7
97	481	KHK-858-896	25.9	1.6	23.5	2.4	34.15	5.07	27.8
98	482	KHK-859-897	42.4	2.4	51.3	4.2	47.048	2.926	46.9
99	483	KHK-860-898-729-571	9.9	1.8	19.1	2.7	13.298	2.656	14.1
100	484	KHK-861-899-730-572	5.0	0.6	3.3	0.4	2.869	0.815	3.7
101	485	KHK-862-900	10.8	1.5	12.2	0.9	12.047	3.121	11.7
102	486	KHK-865	6.3	0.7	7.6	0.5	5.2	0.828	6.4
103	487	KHK-880	11.8	0.8	11.5	1.6	15.705	2.293	13.0
104	488	KHK-882-920	6.5	0.8	6.5	0.9	4.934	1.748	6.0
105	489	KHK-883-921	6.0	1.0	5.3	1.1	5.884	1.209	5.7
106	490	KHK-884-922	11.6	1.4	13.2	1.4	8.24	2.55	11.0
107	491	KHK-885-923	7.9	0.7	9.9	1.0	7.119	2.941	8.3
108	492	KHK-886-924	10.7	7.4	10.9	7.0	4.814	1.257	8.8
109	493	KHK-887-925	14.2	0.7	16.2	1.1	22.141	2.292	17.5
110	494	KHK-888-926	13.3	1.1	17.9	1.4	17.895	1.465	16.4
111	495	KHK-889-927	11.5	0.8	12.3	0.8	13.253	2.142	12.3
112	496	KHK-890-928	8.5	0.7	7.1	0.9	9.332	1.26	8.3
113	497	KHK-891-929	9.0	1.6	8.1	1.3	10.279	2.136	9.1
114	498	KHK-892-930	6.0	1.0	9.4	1.2	7.1	2.124	7.5
115	499	KHK-893-931	11.1	1.0	12.6	1.2	8.773	1.823	10.8
116	500	KHK-894-932	16.1	2.3	19.0	1.7	8.757	1.518	14.6
117	501	KHK-895-933	9.2	0.5	7.4	0.4	14.47	1.915	10.3
118	502	KHK-896-934	10.4	1.1	10.5	1.5	19.901	4.905	13.6
119	503	KHK-897-935	13.1	0.8	14.2	0.9	24.106	2.363	17.1
120	504	KHK-898-936	28.7	1.4	32.0	2.9	25.261	2.143	28.7
121	505	KHK-899-937	10.3	0.8	10.6	1.1	13.265	2.804	11.4
122	506	KHK-900-938	9.2	0.9	10.4	0.8	8.036	1.468	9.2
123	507	KHK-901-939	10.7	2.1	15.3	3.3	9.39	2.386	11.8
124	508	KHK-902-940	17.6	1.2	17.4	2.2	13.271	2.387	16.1
125	509	KHK-903-941	11.5	0.8	15.2	1.2	6.246	1.494	11.0
126	510	KHK-904-942	16.1	3.0	20.6	2.5	12.783	3.392	16.5
127	511	KHK-905-943	10.7	1.7	20.3	0.9	12.273	1.446	14.4
128	512	KHK-906-944	13.2	2.9	24.9	2.4	6.582	2.052	14.9
129	513	KHK-907-945	18.6	11.2	54.3	31.6	14.417	4.546	29.1
130	514	KHK-908-946-777-619	26.7	14.6	46.4	21.7	125.267	92.567	66.1
131	515	KHK-909-947-778-620	7.0	2.5	12.9	4.2	10.062	3.153	10.0
132	516	KHK-910-948-779-621	19.9	3.0	19.2	2.6	19.331	4.315	19.5
133	517	KHK-911-949-780-622	14.0	1.3	12.6	1.0	13.083	3.238	13.2
134	518	KHK-912-950-781-623	20.4	1.1	18.3	1.3	21.258	3.237	20.0
135	519	KHK-913-951-782-624	23.3	0.9	23.6	1.2	28.036	3.803	25.0
136	520	KHK-914-952-783-625	70.2	6.9	69.6	6.1	65.824	7.388	68.5
137	521	KHK-939-977	26.7	4.5	28.7	3.9	22.079	3.952	25.8
138	522	KHK-940-978	14.8	2.5	18.1	3.2	11.124	3.4	14.7
139	523	KHK-941-979	31.0	2.1	35.9	2.4	26.216	2.217	31.1
140	524	KHK-942-980	106.5	6.3	93.3	6.2	55.665	9.171	85.2
141	525	KHK-943-981	15.6	1.6	14.3	1.3	14.943	2.853	14.9
142	526	KHK-944-982	68.2	2.2	64.0	3.2	71.816	5.94	68.0
143	527	KHK-945-983	44.6	1.4	42.6	2.1	60.527	7.442	49.2
144	528	KHK-946-984	42.2	2.1	43.0	2.0	44.669	6.993	43.3
145	529	KHK-947-985	21.0	1.1	22.3	1.9	18.992	1.862	20.8
146	530	KHK-948-986-817	12.7	1.7	13.3	2.9	11.454	2.967	12.5
147	531	KHK-949-987-818	9.1	0.5	9.8	0.8	16.011	13.037	11.7
148	532	KHK-950-988-819	9.6	1.2	8.5	0.8	12.475	3.586	10.2
149	533	KHK-951-989-820	23.5	2.2	25.3	1.9	23.247	6.755	24.0
150	534	KHK-952-990-821	14.5	0.8	18.3	1.6	15.126	4.553	16.0
151	535	KHK-953-991-822	23.0	1.5	25.9	2.2	25.674	5.773	24.9
152	536	KHK-954-992-823	14.1	1.5	16.9	2.7	17.22	2.41	16.1
153	537	KHK-955-993-824	13.0	2.4	14.4	2.5	10.614	3.054	12.7
154	538	KHK-956-994	11.8	2.7	11.9	3.0	17.221	5.593	13.6
155	539	KHK-957-995	6.1	2.1	10.2	1.7	14.207	1.553	10.2
156	540	KHK-958-996	17.4	3.2	17.9	1.0	26.913	3.733	20.7
157	541	KHK-978-1016	43.0	3.9	50.1	4.1	41.716	3.912	45.0
158	542	KHK-982-1020	51.8	2.6	69.1	5.3	67.745	3.674	62.9
159	543	KHK-983-1021	57.0	6.9	60.3	7.5	60.141	8.62	59.2
160	544	KHK-984-1022	15.4	2.1	16.8	2.1	17.339	3.931	16.5
161	545	KHK-985-1023	17.4	2.3	22.7	7.7	16.976	8.222	19.0
162	546	KHK-991-1029	21.4	2.9	22.7	3.9	27.54	9.397	23.9
163	547	KHK-992-1030	11.2	1.3	11.7	1.4	18.836	2.724	13.9
164	548	KHK-993-1031	32.3	5.5	26.5	4.5	19.635	5.352	26.1
165	549	KHK-999-1037	20.8	1.8	24.4	1.8	26.652	2.248	24.0
166	550	KHK-1000-1038	15.3	0.8	20.9	2.4	15.57	2.369	17.3
167	551	KHK-1019-1057	29.8	4.4	29.5	4.4	36.461	4.748	31.9
168	552	KHK-1054-1092	17.3	2.7	14.9	1.9	19.554	3.963	17.2
169	553	KHK-1055-1093	21.0	1.7	22.2	2.4	20.314	4.35	21.2
170	554	KHK-1057-1095	8.8	1.5	8.0	1.4	12.267	1.962	9.7
171	555	KHK-1058-1096	51.6	3.0	52.5	4.1	54.022	5.889	52.7
172	556	KHK-1059-1097	22.0	2.3	13.9	1.9	23.814	3.884	19.9
173	557	KHK-1060-1098	14.7	0.7	10.4	0.8	26.531	6.527	17.2
174	558	KHK-1061-1099	28.3	1.7	22.1	1.5	40.784	3.797	30.4
175	559	KHK-1062-1100	11.6	0.7	12.0	1.1	20.499	2.193	14.7
176	560	KHK-1063-1101	12.6	0.8	9.8	1.2	15.613	1.439	12.7
177	561	KHK-1064-1102	12.7	2.2	9.7	2.5	18.761	3.098	13.7
178	562	KHK-1065-1103	15.2	2.3	13.0	2.0	29.056	2.117	19.1
179	563	KHK-1066-1104	14.0	1.4	13.8	2.9	16.23	2.914	14.7
180	564	KHK-1067-1105	13.7	1.2	10.0	1.2	8.155	3.486	10.6
181	565	KHK-1068-1106	14.0	0.8	11.4	1.1	28.373	4.283	17.9
182	566	KHK-1069-1107	11.8	0.6	9.8	0.7	17.654	3.053	13.1
183	567	KHK-1070-1108	20.2	2.0	14.4	2.7	30.393	6.07	21.7
184	568	KHK-1071-1109	30.0	3.2	22.0	1.5	39.098	8.12	30.4
185	569	KHK-1072-1110	14.5	0.7	12.9	0.8	41.196	15.367	22.8
186	570	KHK-1073-1111	17.7	1.3	14.6	1.7	23.629	3.262	18.6
187	571	KHK-1074-1112-943-785	25.1	2.1	23.0	2.4	34.784	6.555	27.6
188	572	KHK-1075-1113-944-786	12.4	2.5	8.4	1.9	9.594	2.863	10.1
189	573	KHK-1076-1114-945-787	9.8	1.0	7.5	1.0	11.44	3.242	9.6
190	574	KHK-1077-1115-946-788	13.8	1.6	11.9	1.4	19.983	4.226	15.2
191	575	KHK-1078-1116-947-789	12.2	1.4	10.2	1.5	19.236	3.613	13.9
192	576	KHK-1079-1117-948-790	23.0	1.6	20.2	1.4	36.758	3.387	26.6
193	577	KHK-1080-1118-949-791	12.4	0.7	10.3	0.9	23.489	2.532	15.4
194	578	KHK-1081-1119-950-792	19.0	2.8	15.4	1.4	35.094	5.069	23.2
195	579	KHK-1082-1120-951-793	51.6	12.7	36.2	7.1	79.445	13.29	55.8
196	580	KHK-1083-1121-952-794	12.4	0.9	7.6	1.6	29.815	4.341	16.6
197	581	KHK-1084-1122-953-795	17.8	1.5	15.4	1.3	34.538	5.134	22.6
198	582	KHK-1085-1123-954-796	20.4	2.5	19.1	2.4	28.082	3.898	22.5
199	583	KHK-1086-1124-955-797	9.8	1.4	9.1	1.9	22.862	5.973	13.9
200	584	KHK-1087-1125-956-798	25.1	2.4	26.2	5.9	60.678	13	37.3
201	585	KHK-1090-1128	15.7	2.5	14.2	4.3	47.765	6.748	25.9
202	586	KHK-1091-1129	17.1	1.5	16.0	1.2	47.935	10.554	27.0
203	587	KHK-1092-1130	59.1	13.1	81.9	15.1	116.084	17.529	85.7
204	588	KHK-1093-1131	68.9	6.5	72.1	6.2	135.298	14.786	92.1
205	589	KHK-1095-1133	39.3	2.2	34.2	2.9	49.369	9.398	41.0
206	590	KHK-1096-1134	54.0	3.8	58.2	5.6	107.545	13.331	73.2
207	591	KHK-1097-1135	26.5	3.4	22.1	3.0	39.738	6.746	29.5
208	592	KHK-1099-1137	19.4	2.2	21.9	2.9	37.312	8.866	26.2
209	593	KHK-1100-1138	31.5	3.9	31.8	4.6	86.882	29.059	50.1
210	594	KHK-1101-1139	60.8	5.1	65.8	3.9	89.898	14.227	72.2
211	595	KHK-1102-1140	29.8	1.9	25.1	2.9	49.962	4.639	34.9
212	596	KHK-1103-1141	9.9	1.3	5.7	0.9	14.357	4.377	10.0
213	597	KHK-1104-1142	7.9	0.5	5.3	0.7	14.498	2.508	9.2
214	598	KHK-1106-1144	12.0	2.1	6.8	2.4	17.615	5.575	12.1
215	599	KHK-1107-1145	7.6	1.5	3.5	1.2	12.097	3.884	7.7
216	600	KHK-1135-1173	9.8	2.7	6.3	2.1	20.368	5.935	12.2
217	601	KHK-1136-1174	55.0	8.2	57.6	8.7	51.67	14.619	54.8
218	602	KHK-1137-1175	6.9	1.5	5.5	0.9	14.082	4.459	8.8
219	603	KHK-1138-1176	3.4	1.2	2.7	1.0	20.618	4.034	8.9
220	604	KHK-1139-1177	14.2	4.2	10.3	2.6	45.243	8.325	23.2
221	605	KHK-1140-1178	22.3	7.0	14.9	4.7	58.511	10.307	31.9
222	606	KHK-1141-1179	16.8	4.6	18.2	4.1	34.609	7.425	23.2
223	607	KHK-1142-1180	10.0	4.1	8.0	2.3	33.362	7.336	17.1
224	608	KHK-1143-1181	13.7	2.2	12.5	4.2	28.204	4.236	18.1
225	609	KHK-1144-1182	11.7	2.1	14.6	4.6	49.769	18.354	25.3
226	610	KHK-1145-1183	19.0	6.3	22.7	6.7	51.464	20.568	31.1
227	611	KHK-1146-1184	16.8	4.7	14.7	5.5	71.937	37.067	34.5
228	612	KHK-1147-1185	9.0	1.6	9.1	1.1	11.22	4.674	9.7
229	613	KHK-1148-1186	8.8	1.7	6.0	1.8	18.481	8.166	11.1
230	614	KHK-1149-1187	20.1	3.1	18.3	2.8	36.481	11.397	24.9
231	615	KHK-1153-1191	24.4	3.7	20.5	3.0	29.434	6.394	24.8
232	616	KHK-1154-1192	11.9	2.5	8.9	2.2	22.093	6.996	14.3
233	617	KHK-1157-1195	14.5	2.2	11.9	2.2	26.024	4.864	17.5
234	618	KHK-1158-1196	9.3	1.4	5.0	1.5	8.547	5.09	7.6
235	619	KHK-1159-1197	7.3	1.3	11.6	1.3	18.971	4.052	12.6
236	620	KHK-1161-1199	9.9	1.0	4.8	0.7	16.083	3.57	10.3
237	621	KHK-1163-1201	13.7	1.3	11.9	1.4	28.564	6.393	18.0
238	622	KHK-1164-1202	14.6	2.9	10.0	2.5	17.654	4.993	14.1
239	623	KHK-1232-1270	12.0	2.4	13.2	2.6	29.824	4.644	18.4
240	624	KHK-1278-1316-1147-989	14.0	2.6	8.5	3.2	16.096	5.812	12.9
241	625	KHK-1279-1317-1148-990	9.4	2.2	8.2	2.0	23.969	10.352	13.9
242	626	KHK-1280-1318-1149-991	4.5	1.4	5.3	1.2	11.007	1.786	6.9
243	627	KHK-1281-1319-1150-992	4.0	1.3	7.4	2.3	30.368	6.698	13.9
244	628	KHK-1282-1320-1151-993	10.0	1.5	9.4	2.9	10.4	2.173	10.0
245	629	KHK-1283-1321	23.7	3.6	25.9	5.1	20.361	6.111	23.3
246	630	KHK-1284-1322	17.5	1.2	15.1	1.9	23.591	3.59	18.8
247	631	KHK-1285-1323	15.8	1.4	19.5	1.2	13.69	2.902	16.3
248	632	KHK-1286-1324	17.7	2.8	18.7	2.8	18.507	3.994	18.3
249	633	KHK-1287-1325	9.2	3.3	7.7	3.0	10.381	3.571	9.1
250	634	KHK-1288-1326	8.4	2.9	3.8	1.7	11.461	3.398	7.9
251	635	KHK-1289-1327	8.4	2.9	6.4	4.6	11.992	3.025	8.9
252	636	KHK-1290-1328	8.7	1.5	6.7	1.2	8.926	3.258	8.1
253	637	KHK-1291-1329	6.9	0.7	3.8	1.0	7.55	2.469	6.1
254	638	KHK-1292-1330	17.7	1.1	12.8	1.2	30.785	7.501	20.5
255	639	KHK-1293-1331	11.6	1.3	9.7	1.6	15.795	2.45	12.4
256	640	KHK-1294-1332	25.7	1.4	34.2	2.2	33.058	7.279	31.0
257	641	KHK-1295-1333	13.1	1.5	12.2	2.1	15.793	3.001	13.7
258	642	KHK-1297-1335	21.1	1.3	21.8	1.5	24.699	4.755	22.5
259	643	KHK-1323-1361	49.4	8.1	57.3	8.3	44.887	11.548	50.5
260	644	KHK-1325-1363	38.7	2.6	32.1	3.3	28.739	4.127	33.2
261	645	KHK-1326-1364	10.7	0.8	8.2	0.9	20.863	2.809	13.3
262	646	KHK-1327-1365	54.1	2.2	52.2	3.4	94.086	19.883	66.8
263	647	KHK-1328-1366	40.5	1.7	45.3	3.8	44.867	7.194	43.5
264	648	KHK-1329-1367	17.2	1.4	14.4	1.0	32.257	2.561	21.3
265	649	KHK-1330-1368	15.6	1.2	18.9	1.9	19.171	3.863	17.9
266	650	KHK-1331-1369	15.4	1.6	12.9	2.0	42.066	5.461	23.4
267	651	KHK-1332-1370	12.0	0.8	8.5	0.9	10.488	3.467	10.3
268	652	KHK-1333-1371	7.2	0.9	3.7	0.9	8.179	2.058	6.3
269	653	KHK-1334-1372	8.7	1.5	6.5	1.0	7.49	0.985	7.6
270	654	KHK-1335-1373	9.7	0.6	5.2	1.1	9.403	1.153	8.1
271	655	KHK-1336-1374	10.0	1.8	5.2	1.4	14.512	2.647	9.9
272	656	KHK-1385-1423	47.5	3.5	49.0	4.8	44.122	7.678	46.9
273	657	KHK-1387-1425	14.4	2.1	15.9	3.0	22.666	4.122	17.7
274	658	KHK-1388-1426	27.0	2.9	28.2	3.3	34.575	4.985	29.9
275	659	KHK-1389-1427	25.1	3.0	29.5	2.8	18.759	3.577	24.5
276	660	KHK-1538-1588	81.1	15.3	73.7	17.3	71.289	16.403	75.3
277	661	KHK-1540-1590	46.8	4.2	35.5	4.6	23.533	8.593	35.3
278	662	KHK-1542-1592	80.6	4.7	89.1	3.3	85.445	7.942	85.0
279	663	KHK-1665-1708	84.8	4.3	86.6	8.9	116.186	21.343	95.8
280	664	KHK-1666-1709	97.0	2.5	99.6	3.9	115.899	10.593	104.2
281	665	KHK-1667-1710	99.5	4.4	109.4	5.7	123.463	8.991	110.8
282	666	KHK-1707-1750	91.1	3.5	107.3	6.3	123.75	26.01	107.4
283	667	KHK-1708-1751	72.5	5.7	85.8	8.2	76.118	10.985	78.1
284	668	KHK-1709-1752	136.2	5.8	114.5	5.9	75.202	14.025	108.7
285	669	KHK-1869-1918	118.1	21.1	111.2	20.6	97.088	18.195	108.8
286	670	KHK-1870-1919	90.2	8.7	83.2	11.2	87.62	22.519	87.0
287	671	KHK-1871-1920	81.7	4.6	80.6	4.6	119.805	13.019	94.1
288	672	KHK-1872-1921	94.5	4.4	87.4	5.7	74.492	13.782	85.4
289	673	KHK-1873-1922	93.2	5.4	90.9	4.6	88.62	13.98	90.9
290	674	KHK-1874-1923	93.2	6.3	90.7	4.9	74.793	8.913	86.2
291	675	KHK-1875-1924	86.5	8.5	76.3	8.2	45.965	7.621	69.6
292	676	KHK-1876-1925	73.7	9.7	61.1	8.7	34.813	6.807	56.6
293	677	KHK-1877-1926	72.7	3.9	51.0	5.3	45.122	9.468	56.3
294	678	KHK-1878-1927	75.8	4.8	79.7	8.0	54.716	8.031	70.1
295	679	KHK-1879-1928	90.6	8.4	77.0	9.0	86.62	13.219	84.7
296	680	KHK-1880-1929	100.4	8.5	84.5	9.8	85.501	13.022	90.1
297	681	KHK-1900-1949	104.9	12.1	96.5	15.3	70.712	8.039	90.7
298	682	KHK-1905-1954	71.4	17.0	71.0	22.1	35.273	14.001	59.2
299	683	KHK-1971-2025	77.5	25.4	128.3	28.3	34.399	9.559	80.1
300	684	KHK-1974-2028	74.2	6.0	64.4	7.7	50.763	6.122	63.1
301	685	KHK-1975-2029	80.8	7.8	80.5	7.1	55.779	10.606	72.3
302	686	KHK-1976-2030	69.2	6.8	73.5	8.5	53.873	16.663	65.5
303	687	KHK-1978-2032	73.7	10.3	73.6	10.0	57.334	7.876	68.2
304	688	KHK-1979-2033	81.2	9.5	95.1	13.6	43.016	8.767	73.1
305	689	KHK-2032-2086	94.2	6.3	124.6	9.4	99.842	10.374	106.2
306	690	KHK-2035-2089	66.5	6.2	72.2	14.5	47.134	6.316	61.9
307	691	KHK-2036-2090	82.4	9.1	110.4	15.5	56.532	12.458	83.1
308	692	KHK-2037-2091	88.9	8.1	72.5	7.9	48.451	10.859	70.0
309	693	KHK-2038-2092	74.2	3.6	58.7	4.7	44.437	4.32	59.1
310	694	KHK-2039-2093	75.1	5.3	76.3	8.1	51.597	6.117	67.7
311	695	KHK-2040-2094	75.9	8.5	70.0	11.2	57.499	6.477	67.8
312	696	KHK-2041-2095	80.5	4.1	78.1	3.5	78.593	10.192	79.1
313	697	KHK-2042-2096	79.1	5.2	84.7	4.0	88.699	8.352	84.2
314	698	KHK-2043-2097	72.0	3.8	70.0	2.7	83.791	8.891	75.3
315	699	KHK-2044-2098	37.6	12.9	32.3	13.4	30.83	15.258	33.6
316	700	KHK-2045-2099	101.3	8.7	87.4	11.3	54.839	12.261	81.2
317	701	KHK-2067-2121	88.2	5.1	78.4	2.6	75.916	8.438	80.8
318	702	KHK-2069-2123	83.1	3.7	84.9	5.2	63.679	10.343	77.2
319	703	KHK-2091-2145	83.8	5.4	87.1	8.9	53.463	10.34	74.8
320	704	KHK-2092-2146	85.4	6.2	89.7	7.8	68.656	5.01	81.3
321	705	KHK-2093-2147	102.7	21.4	65.4	19.9	71.693	14.857	79.9
322	706	KHK-2094-2148	88.8	6.4	94.1	6.7	53.85	10.392	78.9
323	707	KHK-2095-2149	76.2	21.4	97.2	25.9	47.372	14.735	73.6
324	708	KHK-2096-2150	87.9	16.4	78.6	13.2	49.454	14.398	72.0
325	709	KHK-2105	92.1	7.7	90.8	11.5	97.683	10.156	93.5
326	710	KHK-2148-2197	86.5	5.8	79.3	16.0	76.198	8.142	80.7
327	711	KHK-2149-2198	71.3	3.7	73.8	4.2	55.558	6.731	66.9
328	712	KHK-2150-2199	92.1	7.1	97.5	4.9	75.703	6.126	88.4
329	713	KHK-2151-2200	96.2	3.7	108.0	10.3	91.908	5.852	98.7
330	714	KHK-2152-2201	78.7	6.5	74.7	9.6	42.766	6.332	65.4
331	715	KHK-2153-2202	95.2	14.8	73.9	13.1	47.169	10.902	72.1
332	716	KHK-2154-2203	114.1	11.9	92.3	7.2	61.728	10.943	89.4
333	717	KHK-2155-2204	92.1	8.4	83.1	3.8	119.537	11.18	98.2
334	718	KHK-2156-2205	104.7	5.4	91.6	4.7	148.445	15.208	114.9
335	719	KHK-2157-2206	94.2	8.4	92.0	10.4	68.735	7.132	85.0
336	720	KHK-2159-2208	85.4	4.5	78.4	6.2	62.397	10.642	75.4
337	721	KHK-2160-2209	72.7	1.9	81.8	5.3	56.483	11.255	70.3
338	722	KHK-2161-2210	93.7	14.6	74.3	10.9	18.252	5.102	62.1
339	723	KHK-2162-2211	106.7	11.3	127.3	17.9	53.455	15.254	95.8
340	724	KHK-2163-2212	79.5	8.2	91.6	6.4	49.199	6.236	73.5
341	725	KHK-2164-2213	101.1	13.4	115.2	20.4	84.893	29.662	100.4
342	726	KHK-2165-2214	97.0	10.8	102.1	9.9	76.079	10.525	91.7
343	727	KHK-2166-2215	91.6	20.4	89.3	22.7	55.353	11.894	78.8
344	728	KHK-2170-2219	75.9	4.5	89.7	4.6	68.461	8.991	78.0
345	729	KHK-2196-2245	60.3	2.9	65.1	4.4	43.35	5.951	56.3
346	730	KHK-2197-2246	85.4	8.4	98.9	9.4	65.81	6.865	83.4
347	731	KHK-2198-2247	89.4	15.0	108.1	10.3	44.371	7.323	80.6
348	732	KHK-2199-2248	97.2	14.8	91.3	16.1	49.493	8.874	79.3
349	733	KHK-2200-2249	104.7	10.7	111.8	13.5	47.327	5.488	87.9
350	734	KHK-2201-2250	100.3	11.7	102.7	13.8	52.984	11.652	85.3
351	735	KHK-2205	96.9	17.0	88.0	10.9	55.021	8.208	80.0
352	736	KHK-2238	89.1	8.0	99.6	8.2	113.917	11.636	100.9
353	737	KHK-2260-2309	111.5	13.2	110.2	11.9	95.452	20.407	105.7
354	738	KHK-2261-2310	103.6	9.1	106.9	12.4	97.581	15.894	102.7
355	739	KHK-2262-2311	141.3	14.6	132.9	11.8	112.052	19.366	128.7
356	740	KHK-2263-2312	104.0	10.5	80.5	11.4	59.852	7.342	81.4
357	741	KHK-2264-2313	100.7	17.1	88.6	15.5	53.023	12.056	80.8
358	742	KHK-2265-2314	103.2	11.2	103.8	11.9	60.929	8.309	89.3
359	743	KHK-2266-2315	119.8	8.9	110.3	8.5	66.846	9.19	99.0
360	744	KHK-2299	77.7	3.2	72.0	5.9	69.804	11.442	73.2
361	745	KHK-2317-2366	81.2	3.0	84.4	7.7	66.04	7.299	77.2
362	746	KHK-2318-2367	86.6	3.7	97.0	5.2	66.519	4.573	83.4
363	747	KHK-2319-2368	127.5	12.5	102.9	8.9	82.338	11.524	104.2
364	748	KHK-2320-2369	98.9	11.1	94.2	15.6	59.154	17.115	84.1
365	749	KHK-2321-2370	127.6	13.8	127.6	16.5	81.979	19.313	112.4
366	750	KHK-2322-2371	83.3	9.1	68.5	8.6	78.252	14.344	76.7
367	751	KHK-2323-2372	83.2	7.3	79.8	7.7	57.202	9.062	73.4
368	752	KHK-2324-2373	95.0	3.0	101.4	5.2	118.194	16.285	104.9
369	753	KHK-2325-2374	123.0	15.6	153.7	23.1	188.498	24.768	155.1
370	754	KHK-2326-2375	94.5	12.0	101.2	8.2	110.056	29.627	101.9
371	755	KHK-2332	96.2	11.9	122.2	10.2	120.096	15.829	112.8
372	756	KHK-2333	97.9	12.7	73.0	9.9	50.041	16.617	73.7
373	757	KHK-2335	104.4	9.0	81.2	7.5	33.77	6.814	73.1
374	758	KHK-2340	64.4	14.3	45.7	12.3	34.661	10.468	48.2
375	759	KHK-2341	61.9	10.6	53.7	6.9	43.579	8.14	53.1
376	760	KHK-2346	104.5	9.2	92.5	12.6	104.386	32.315	100.4
377	761	KHK-2352	78.5	6.7	83.6	8.0	97.544	18.168	86.6
378	762	KHK-2358	78.8	7.0	75.7	9.9	64.974	13.327	73.1
379	763	KHK-2359	89.5	9.0	91.8	8.2	71.314	7.358	84.2
380	764	KHK-2360	132.4	26.2	82.8	8.7	159.973	15.729	125.0
381	765	KHK-2361	110.4	8.5	87.7	7.8	87.634	7.799	95.3
382	766	KHK-2362	92.5	12.8	72.7	7.2	84.427	12.833	83.2
383	767	KHK-2363	100.4	9.2	76.1	14.4	52.066	18.068	76.2
384	768	KHK-2364	116.4	9.8	88.5	8.3	120.931	26.855	108.6
385	769	KHK-2365	100.5	5.7	92.9	5.2	132.668	7.14	108.7
386	770	KHK-2366	129.3	7.2	113.5	5.7	135.163	16.2	126.0
387	771	KHK-2367	123.3	9.0	105.9	9.2	136.356	9.025	121.9

Example 3: RNAi Oligonucleotide Inhibition of Both KHK Isoforms In Vivo

The in vitro screening assay in Example 2 validated the ability of KHK DsiRNA to knock-down both isoforms of KHK (KHK-All). To confirm the ability of the RNAi oligonucleotides to knockdown both KHK-A and KHK-C isoforms, a side-by-side HDI mouse model was used. First, the nucleotide sequences comprising a subset of the 384 DsiRNAs identified in Example 2, and that recognize human/NHP-conserved KHK, were used to generate corresponding double-stranded RNAi oligonucleotides comprising a nicked tetraloop GalNAc-conjugated structure (referred to herein as “GalNAc-conjugated KHK oligonucleotides” or “GalNAc-KHK constructs”) having a 36-mer passenger strand and a 22-mer guide strand (Table 3). Specifically, to generate the 22-mer guide strand, the 19-mer core antisense strand sequences used in Example 2 (e.g., SEQ ID NOs: 948-953) were modified to have a phosphorylated uracil at the 5′ end and two guanines at the 3′ end. To generate the 36-mer passenger strand, an adenine corresponding to the phosphorylated uracil in the antisense strand and a 16-mer stem loop (SEQ ID NO: 871) were added to the 3′ end of the 19-mer core sense strand sequences used in Example 2 (e.g., SEQ ID NOs: 942-947). Further, the nucleotide sequences comprising the passenger strand and guide strand of the GalNAc-conjugated KHK oligonucleotides have a distinct pattern of modified nucleotides and phosphorothioate linkages (e.g., see FIG. 2A, FIG. 2B and Table 3 for schematics of the generic structure and key of chemical modifications; referred to herein as Low-2′-Fluoro (3PS) and Low-2′-Fluoro (2PS), respectively, together as the Low-2′-Fluoro pattern for GalNAc-conjugated KHK oligonucleotides). The three adenosine nucleotides comprising the tetraloop are each conjugated to a GalNAc moiety (CAS #: 14131-60-3). The modification pattern is represented below in two interchangeable modification keys.

Low-2′-Fluoro (3PS) Modification Pattern for
GalNAc-KHK Constructs (5′ Antisense 3PS)
Sense Strand:
5′-mX-S-mX-mX-mX-mX-mX-mX-fX-fX-fX-fX-mX-mX-
mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-
[ademA-GalNAc]- [ademA-GalNAc]- [ademA-
GalNAc]-mX-mX-mX-mX-mX-mX-3′.
Hybridized to:
Antisense Strand:
5′-[MePhosphonate-4O-mX]-S-fX-S-fX-S-fX-fX-
mX-fX-mX-mX-fX-mX-mX-mX-fX-mX-mX-mX-mX-mX-
mX-S-mX-S-mX-3′
(Modification key: Table 3).
Or, represented as:
Sense Strand:
[mXs][mX][mX][mX][mX][mX][mX][fX][fX][fX]
[fX][mX][mX][mX][mX][mX][mX][mX][mX][mX]
[mX][mX][mX][mX][mX][mX][mX][ademA-GalNAc]
[ademA-GalNAc][ademA-GalNAc][mX][mX][mX]
[mX][mX][mX]
Hybridized to:
Antisense Strand:
[MePhosphonate-4O-mXs][fXs][fXs][fX][fX]
[mX][fX][mX][mX][fX][mX][mX][mX][fX][mX]
[mX][mX][mX][mX][mXs][mXs][mX]
(Modification key: Table 3).
Low-2′Fluoro (2PS) Modification Pattern
for GalNAc-KHK Constructs (5′ Antisense
2PS)
Sense Strand:
5′-mX-S-mX-mX-mX-mX-mX-mX-fX-fX-fX-fX-mX-
mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-
mX-mX-[ademA-GalNAc]- [ademA-GalNAc]-
[ademA-GalNAc]-mX-mX-mX-mX-mX-mX-3′.
Hybridized to:
Antisense Strand:
5′-[MePhosphonate-4O-mX]-S-fX-S-fX-fX-fX-
mX-fX-mX-mX-fX-mX-mX-mX-fX-mX-mX-mX-mX-
mX-mX-S-mX-S-mX-3′
(Modification key: Table 3).
Or, represented as:
Sense Strand:
[mXs][mX][mX][mX][mX][mX][mX][fX][fX][fX]
[fX][mX][mX][mX][mX][mX][mX][mX][mX][mX]
[mX][mX][mX][mX][mX][mX][mX][ademA-GalNAc]
[ademA-GalNAc][ademA-GalNAc][mX][mX][mX]
[mX][mX][mX]
Hybridized to:
Antisense Strand:
[MePhosphonate-4O-mXs][fXs][fX][fX][fX]
[mX][fX][mX][mX][fX][mX][mX][mX][fX][mX]
[mX][mX][mX][mX][mXs][mXs][mX]

TABLE 3
Modification Key
Symbol          Modification/linkage
Key 1
mX              2′-O-methyl modified nucleotide
fX              2′-fluoro modified nucleotide
-S-             phosphorothioate linkage
—               phosphodiester linkage
[MePhosphonate- 5′-methoxyphosphonate-4′-oxy modified nucleotide
4O-mX]
ademA-GalNAc    GalNAc attached to an adenine nucleotide
Key 2
[mXs]           2′-O-methyl modified nucleotide with a
                phosphorothioate linkage to the neighboring nucleotide
[fXs]           2′-fluoro modified nucleotide with a
                phosphorothioate linkage to the neighboring nucleotide
[mX]            2′-O-methyl modified nucleotide with phosphodiester
                linkages to neighboring nucleotides
[fX]            2′-fluoro modified nucleotide with phosphodiester
                linkages to neighboring nucleotides

The GalNAc-KHK constructs were then used to evaluate inhibition efficacy in mice. Specifically, 6-8-week-old female CD-1 mice (n=5) were subcutaneously administered the indicated GalNAc-conjugated KHK oligonucleotides (Table 4) at a dose of 2 mg/kg formulated in PBS. A control group of mice (n=5) were administered only PBS. Three days later (72 hours), the mice were hydrodynamically injected (HDI) either with a DNA plasmid (pCMV6-KHK-C, Cat #: RC223488, OriGene) encoding the full human KHK gene (NM_006488.3) (25 μg) or plasmid (pCMV6-KHK-A, Cat #; RC202424, OriGene) encoding the full human KHK-A gene (NM_000221) under control of a ubiquitous cytomegalovirus (CMV) promoter sequence. One day after introduction of the DNA plasmid, liver samples from HDI mice were collected. The values were normalized for transfection efficiency using the NeoR gene included on the DNA plasmid.

Total RNA isolated from mouse livers were used to assess relative KHK mRNA expressions by qRT-PCR. The TaqMan RT-qPCR probes from Life Technologies were used to evaluate [3′ assay (Forward-1026; TGGAGGTGGAGAAGCCA (SEQ ID NO: 865), Reverse-1157; GACCATACAAGCCCCTCAAG (SEQ ID NO:866), Probe-1080; TGGTGTTTGTCAGCAAAGATGTGGC (SEQ ID NO:867)) and a 5′ assay (Forward-496; AGGAAGCTCTGGGAGTA (SEQ ID NO: 868), Reverse-596; CCTCCTTAGGGTACTTGTC (SEQ ID NO: 869), Probe-518; ATGGAAGAGAAGCAGATCCTGTGCG (SEQ ID NO: 870))]. The values were normalized for transfection efficiency using the NeoR gene included on the DNA plasmid. HDI mice were generated as described above but using a human KHK-A plasmid or a human KHK-C plasmid. The mice were treated in groups of 5 with the GalNAc-KHK constructs in Table 4 (with the Low-2′-Fluoro modification pattern). Livers were collected and mRNA measured using primer pairs recognizing KHK-All, KHK-C, or KHK-A. The results confirmed that GalNAc-KHK constructs designed to target all KHK transcripts demonstrate successful knockdown in both the human KHK-A and KHK-C HDI mouse models (FIG. 3).

TABLE 4
GalNAc-KHK Constructs Evaluated In
KHK-C and KHK-A HDI Mouse Models
	Sense	Anti-sense	Sense	Anti-sense
	Strand	Strand	Strand	Strand
	Unmodified	Unmodified	Modified	Modified
Name	SEQ ID NO	SEQ ID NO	SEQ ID NO	SEQ ID NO
KHK-516	39	423	775	820
KHK-865	102	486	779	824
KHK-882	104	488	780	825
KHK-885	107	491	782	827
KHK-1078	191	575	785	830
KHK-1334	269	653	804	849

Example 4: Changes in Modification Pattern of KHK-Targeting RNAi Oligonucleotides Maintains mRNA Inhibition Efficacy

To assess whether modification patterns may impact the targeting efficiency and stability of GalNAc-KHK constructs, two unique patterns were analyzed in HDI mice. Specifically, the modification patterns used were the Low-2′-fluoro pattern described in Example 3 (see FIG. 2A and FIG. 2B) and a Med-2′-fluoro pattern (see FIG. 4A).

Med-2′-Fluoro Modification Pattern for GalNAc-
KHK Constructs
Sense Strand:
5′-mX-S-mX-fX-mX-mX-mX-mX-fX-fX-fX-mX-fX-fX-
mX-mX-mX-fX-mX-mX-mX-mX-mX-mX-mX-mX-mX-mX-
[ademA-GalNAc]- [ademA-GalNAc]- [ademA-
GalNAc]-mX-mX-mX-mX-mX-mX-3′.
Hybridized to:
Antisense Strand:
5′-[MePhosphonate-4O-mX]-S-fX-S-fX-S-fX-fX-
mX-fX-fX-mX-fX-mX-mX-mX-fX-mX-fX-mX-mX-fX-
mX-S-mX-S-mX-3′
(Modification key: Table 3).
Or, represented as:
Sense Strand:
[mXs][mX][fX][mX][mX][mX][mX][fX][fX][fX]
[mX][fX][fX][mX][mX][mX][fX][mX][mX][mX]
[mX][mX][mX][mX][mX][mX][mX][ademA-GalNAc]
[ademA-GalNAc][ademA-GalNAc][mX][mX][mX]
[mX][mX][mX]
Hybridized to:
Antisense Strand:
[MePhosphonate-4O-mXs][fXs][fXs][fX][fX]
[mX][fX][fX][mX][fX][mX][mX][mX][fX][mX]
[fX][mX][mX][fX][mXs][mXs][mX]
(Modification key: Table 3).

HDI mice were generated as described in Example 3. Mice were treated with Low-2′-Fluoro or Med-2′-Fluoro modified KHK constructs (Table 5). 72 hours after treatment, mice were hydrodynamically injected with [pcDNA3.1-KHK-C, encoding the full human KHK gene (NM_006488)]. Livers were collected and processed as described in Example 3. A group of GalNAc-KHK constructs (KHK-0861, -0865, -0882, -0883, -0885) were mixed together and used as a positive control for inhibition. Both modification patterns resulted in inhibition of KHK mRNA in mice (FIG. 4B-4E). These results demonstrate that both modification patterns provided knockdown of the target mRNA.

TABLE 5
GalNAc-KHK Constructs for Modification Pattern Assay
		Sense	Anti-sense	Sense	Anti-sense
		Strand	Strand	Strand	Strand
	Modifi-	Unmodi-	Unmodi-	Modified	Modified
	cation	fied SEQ	fied SEQ	SEQ	SEQ
Name	pattern	ID NO	ID NO	ID NO	ID NO
KHK-861	Low-2′F	100	484	778	823
KHK-861	Med-2′F	100	484	808	853
KHK-865	Low-2′F	102	486	779	824
KHK-865	Med-2′F	102	486	809	854
KHK-882	Low-2′F	104	488	780	820
KHK-882	Med-2′F	104	488	810	855
KHK-883	Low-2′F	105	489	781	826
KHK-883	Med-2′F	105	489	811	856
KHK-885	Low-2′F	107	491	782	827
KHK-885	Med-2′F	107	491	812	857
KHK-1288	Low-2′F	250	634	787	832
KHK-1288	Med-2′F	250	634	816	861
KHK-1290	Low-2′F	252	636	788	833
KHK-1290	Med-2′F	252	636	817	862
KHK-1334	Low-2′F	269	653	804	849
KHK-1334	Med-2′F	269	653	818	863
KHK-516	Med-2′F	39	423	805	850
KHK-804	Med-2′F	N/A	N/A	806	851
KHK-829	Med-2′F	92	476	807	852
KHK-1076	Med-2′F	189	573	814	859
KHK-1078	Med-2′F	191	575	815	860

Example 5: RNAi Oligonucleotide Inhibition of KHK Expression In Vivo Mouse HDI KHK Knockdown Screening Studies

The GalNAc-conjugated KHK oligonucleotides listed in Table 6 were evaluated in HDI mice as described in Example 3. GalNAc-KHK construct treatment effectively reduced KHK-All mRNA (FIG. 5). When using primers specific for the KHK-C isoform, the GalNAc-KHK constructs were still effective at reducing mRNA (FIG. 5).

TABLE 6
GalNAc-KHK Constructs Assayed in HDI Model
		Sense Strand	Antisense Strand
	Name	Modified SEQ ID NO	Modified SEQ ID NO
	KHK-885	782	827
	KHK-869	795	840
	KHK-873	796	841
	KHK-879	797	842
	KHK-881	798	843
	KHK-896	799	844
	KHK-1064	800	845
	KHK-1075	784	829
	KHK-1077	801	846
	KHK-1080	802	847
	KHK-1106	803	848
	KHK-1147	794	839
	KHK-1148	789	834
	KHK-1152	790	835
	KHK-1154	791	836
	KHK-1155	792	837
	KHK-1277	793	838

Additional constructs (Table 7) were assayed using the same methods and found effective knock-down for KHK-All and KHK-C (FIGS. 6A and 6B). Similarly, endogenous mouse KHK was reduced by GalNAc-KHK constructs which align with mouse KHK mRNA (FIG. 6C). Overall, both HDI studies identified GalNAc-KHK constructs effective at reducing KHK mRNA in vivo.

TABLE 7
GalNAc-KHK Constructs Assayed in HDI Model
		Sense Strand	Antisense Strand
	Name	Modified SEQ ID NO	Modified SEQ ID NO
	KHK-1054	783	828
	KHK-510	774	819
	KHK-516	775	820
	KHK-829	776	821
	KHK-860	777	822
	KHK-861	778	823
	KHK-865	779	824
	KHK-882	780	825
	KHK-883	781	826
	KHK-885	782	827
	KHK-1075	784	829
	KHK-1078	785	830
	KHK-1281	786	831
	KHK-1288	787	832
	KHK-1290	788	833
	KHK-1334	804	849

Example 6: RNAi Oligonucleotide Inhibition of KHK Expression and Studies in Non-Human Primates

Single-dose Non-Human Primate (NHP) Studies

Effective GalNAc-KHK constructs identified in the HDI mouse studies were assayed for targeting efficiency in non-human primates. Specifically, GalNAc-conjugated KHK oligonucleotides listed in Table 8 were evaluated in non-naïve cynomolgus monkeys (Macaca fascicularis). In this study, the monkeys were grouped so that their mean body weights (about 5.4 kg) were comparable between the control and experimental groups. Each cohort contained at least two female and at least two male subjects. The GalNAc-conjugated KHK oligonucleotides were administered subcutaneously at a dose of 6 mg/kg on Study Day 0. Blood samples were collected one week prior to dosing (Day −7), on the dosing date (Day 0) and days 28, 56 and 84 after dosing. Ultrasound-guided core needle liver biopsies were collected on Study Days −7, 28, 56 and 84. At each time point, total RNA derived from the liver biopsy samples was subjected to qRT-PCR analysis to measure KHK mRNA in oligonucleotide-treated monkeys relative to those treated with a comparable volume of PBS. To normalize the data, the measurements were made relative to the geometric mean of two reference genes, PPIB and 18S rRNA. The following TaqMan qPCR probes purchased from Life Technologies, Inc, were used to evaluate gene expressions: Forward—TGCCTTCATGGGCTCAATG (SEQ ID NO: 772); Reverse—TCGGCCACCAGGAAGTCA (SEQ ID NO: 773); Fam probe-CCCTGGCCATGTTG (SEQ ID NO:864)). As shown in FIG. 7A (Day 28), treating NHPs with the GalNAc-conjugated KHK oligonucleotides listed in Table 8 inhibited KHK expression in the liver, as determined by a reduced amount of KHK mRNA in liver samples from oligonucleotide-treated NHPs relative to NHPs treated with PBS. The mean percent reduction of KHK mRNA in the liver samples of treated NHPs is indicated above the set of data points for each treatment group. Days 56 and 84 were also measured (FIGS. 7B and 7C) and a plot of the mean values over each time point is shown in FIG. 7D. For all time points evaluated, almost all the tested GalNAc-conjugated KHK oligonucleotides significantly inhibited KHK mRNA expression. In the same samples, KHK protein levels were detected using rabbit anti-Ketohexokinase (Abcam, AB197593) and anti-rabbit Detection Module for Sally Sue (Protein Simple, cat #DM-001). As shown in FIGS. 8A-8C, at the 28-day timepoint, GalNAc-KHK constructs inhibit KHK protein expression, as normalized to the vinculin control and slowly increases by Day 86. These results demonstrate that treating NHPs with the GalNAc-conjugated KHK oligonucleotides reduces the amount of KHK mRNA in the liver and concomitantly reduces the amount of KHK protein in the liver. However, this correlation is reduced over time after the initial dose (FIGS. 9A-9C).

Taken together, these results show that GalNAc-conjugated KHK oligonucleotides designed to target human total KHK mRNA inhibit total KHK expression in vivo (as determined by the reduction of the amount of KHK mRNA and protein).

TABLE 8
Single-dose GalNAc-KHK Constructs for NHP Study
Name	Sense strand SEQ ID NO	Anti-sense strand SEQ ID NO
KHK-516	775	820
KHK-865	779	824
KHK-882	780	825
KHK-885	782	827
KHK-1078	785	830
KHK-1334	804	849

SEQUENCE LISTING
	Descrip-
	tion
	Species
	(Hs-Mf-			SEQ
	Ms-Rn			ID
Name	(rat))	Strand	Sequence	NO
Human	Human	N/A	AGGCAGGGCTGCAGATGCGAGGCCCAGC	1
(Hs) KHK	(Hs)		TGTACCTCGCGTGTCCCGGGTCGGGAGTC
nucleotide			GGAGACGCAGGTGCAGGAGAGTGCGGGG
sequence			CAAGTAGCGCATTTTCTCTTTGCATTCTCG
NM_00648			AGATCGCTTAGCCGCGCTTTAAAAAGGTTT
8.3			GCATCAGCTGTGAGTCCATCTGACAAGCG
			AGGAAACTAAGGCTGAGAAGTGGGAGGC
			GTTGCCATCTGCAGGCCCAGGCAACCTGC
			TACGGGAAGACCGGGGACCAAGACCTCT
			GGGTTGGCTTTCCTAGACCCGCTCGGGTC
			TTCGGGTGTCGCGAGGAAGGGCCCTGCT
			CCTTTCGTTCCCTGCACCCCTGGCCGCTG
			CAGGTGGCTCCCTGGAGGAGGAGCTCCC
			ACGCGGAGGAGGAGCCAGGGCAGCTGGG
			AGCGGGGACACCATCCTCCTGGATAAGAG
			GCAGAGGCCGGGAGGAACCCCGTCAGCC
			GGGCGGGCAGGAAGCTCTGGGAGTAGCC
			TCATGGAAGAGAAGCAGATCCTGTGCGTG
			GGGCTAGTGGTGCTGGACGTCATCAGCCT
			GGTGGACAAGTACCCTAAGGAGGACTCGG
			AGATAAGGTGTTTGTCCCAGAGATGGCAG
			CGCGGAGGCAACGCGTCCAACTCCTGCA
			CCGTTCTCTCCCTGCTCGGAGCCCCCTGT
			GCCTTCATGGGCTCAATGGCTCCTGGCCA
			TGTTGCTGACTTCCTGGTGGCCGACTTCA
			GGCGGCGGGGCGTGGACGTGTCTCAGGT
			GGCCTGGCAGAGCAAGGGGGACACCCCC
			AGCTCCTGCTGCATCATCAACAACTCCAAT
			GGCAACCGTACCATTGTGCTCCATGACAC
			GAGCCTGCCAGATGTGTCTGCTACAGACT
			TTGAGAAGGTTGATCTGACCCAGTTCAAGT
			GGATCCACATTGAGGGCCGGAACGCATCG
			GAGCAGGTGAAGATGCTGCAGCGGATAGA
			CGCACACAACACCAGGCAGCCTCCAGAGC
			AGAAGATCCGGGTGTCCGTGGAGGTGGA
			GAAGCCACGAGAGGAGCTCTTCCAGCTGT
			TTGGCTACGGAGACGTGGTGTTTGTCAGC
			AAAGATGTGGCCAAGCACTTGGGGTTCCA
			GTCAGCAGAGGAAGCCTTGAGGGGCTTGT
			ATGGTCGTGTGAGGAAAGGGGCTGTGCTT
			GTCTGTGCCTGGGCTGAGGAGGGCGCCG
			ACGCCCTGGGCCCTGATGGCAAATTGCTC
			CACTCGGATGCTTTCCCGCCACCCCGCGT
			GGTGGATACACTGGGAGCTGGAGACACCT
			TCAATGCCTCCGTCATCTTCAGCCTCTCCC
			AGGGGAGGAGCGTGCAGGAAGCACTGAG
			ATTCGGGTGCCAGGTGGCCGGCAAGAAG
			TGTGGCCTGCAGGGCTTTGATGGCATCGT
			GTGAGAGCAGGTGCCGGCTCCTCACACAC
			CATGGAGACTACCATTGCGGCTGCATCGC
			CTTCTCCCCTCCATCCAGCCTGGCGTCCA
			GGTTGCCCTGTTCAGGGGACAGATGCAAG
			CTGTGGGGAGGACTCTGCCTGTGTCCTGT
			GTTCCCCACAGGGAGAGGCTCTGGGGGG
			ATGGCTGGGGGATGCAGAGCCTCAGAGC
			AAATAAATCTTCCTCAGAGCCAGCTTCTCC
			TCTCAATGTCTGAACTGCTCTGGCTGGGC
			ATTCCTGAGGCTCTGACTCTTCGATCCTCC
			CTCTTTGTGTCCATTCCCCAAATTAACCTC
			TCCGCCCAGGCCCAGAGGAGGGGCTGCC
			TGGGCTAGAGCAGCGAGAAGTGCCCTGG
			GCTTGCCACCAGCTCTGCCCTGGCTGGG
			GAGGACACTCGGTGCCCCACACCCAGTGA
			ACCTGCCAAAGAAACCGTGAGAGCTCTTC
			GGGGCCCTGCGTTGTGCAGACTCTATTCC
			CACAGCTCAGAAGCTGGGAGTCCACACCG
			CTGAGCTGAACTGACAGGCCAGTGGGGG
			GCAGGGGTGCGCCTCCTCTGCCCTGCCC
			ACCAGCCTGTGATTTGATGGGGTCTTCATT
			GTCCAGAAATACCTCCTCCCGCTGACTGC
			CCCAGAGCCTGAAAGTCTCACCCTTGGAG
			CCCACCTTGGAATTAAGGGCGTGCCTCAG
			CCACAAATGTGACCCAGGATACAGAGTGT
			TGCTGTCCTCAGGGAGGTCCGATCTGGAA
			CACATATTGGAATTGGGGCCAACTCCAATA
			TAGGGTGGGTAAGGCCTTATAATGTAAAG
			AGCATATAATGTAAAGGGCTTTAGAGTGAG
			ACAGACCTGGATTAAAATCTGCCATTTAAT
			TAGCTGCATATCACCTTAGGGTACAGCACT
			TAACGCAATCTGCCTCAATTTCTTCATCTG
			TCAAATGGAACCAATTCTGCTTGGCTACAG
			AATTATTGTGAGGATAAAATCATATATAAAA
			TGCCCAGCATGATGCCTGATGTGTA
Cyno-	Cyno-	N/A	GGGGCCGGGCAGCCGCGACCACGGTCTT	2
molgus	molgus		CAGGCAGGGCTGCAGATGCAGGCCCAGC
monkey	monkey		TCTACCTCGCGGGTCCAGGGTCGGGAGT
(Mf)	(Mf)		CCGAGACGCAGGTGCAGCAGAGGGCGGG
KHK			GCACGTAGCGCATTTCCAGCGCATTTTCT
nucleotide			CTTTGCATTCTCGAGATCGCTTAGCCGCG
sequence			CTTTAGAAGGGTTTGCATCAGCTCCGAGT
XM_00557			CCATCTGACAAGCGAGGAAACTGAGGCTG
6322.2			AGAAGTGGGAGGCGTTGCCATCTGCAGG
			CCCAGGCAACCTGCTACGGGAAGACCGG
			GGGCCAAGACCTCCGGGTTGGCTTTCCCA
			GGCCAGCTTGGGTCTTCGGGTGTCGGGA
			GCAAAGGCCCAGCTCCTTTCGTTTCCTGC
			ACCCCTCGCCGCTGCAGGTGGCTCCCCG
			GAGGAGGAGCTCCCACGCGGAGGAGGAG
			CCAGGGCAGCTGGGAGCGAGGACACCAT
			CCTCCTGGATAACAGGCAGAGGCCGGGA
			GGAACCCGTCAGTCGGGCGGGCAGGAAG
			CTCTGGGATCAGCCTCATGGAAGAGAAGC
			AGATCCTGTGCGTGGGGCTAGTGGTGCTG
			GACGTCATCAGCCTGGTGGACAAGTACCC
			TAAGGAGGACTCAGAGATAAGGTGCTTGT
			CCCAGAGATGGCAACGCGGAGGCAACGC
			GTCCAACTCCTGCACCGTTCTCTCCCTGC
			TCGGAGCCCCCTGTGCCTTCATGGGCTCA
			ATGGCCCCTGGCCATGTTGCTGACTTCCT
			GGTGGCCGACTTCAGGCGGCGGGGTGTG
			GACGTGTCTCAGGTGGCCTGGCAGAGCAA
			GGGGGACACCCCCAGCTCCTGCTGCATCA
			TCAACAACTCCAATGGCAACCGTACCATTG
			TGCTCCATGACACGAGCCTGCCAGATGTG
			TCTGCTACGGACTTTGAGAAGGTTGATCT
			GACCCAGTTCAAGTGGATCCACATTGAGG
			GCCGGAATGCATCGGAGCAGGTGAAGAT
			GCTGCAGCGGATAGACGCGCACAACACCA
			GGCAGCCTCCAGAGCAGAAGATCCGGGT
			GTCCGTGGAGGTGGAGAAGCCACAAGAG
			GAGCTCTTTCAGCTGTTTGGCTACGGAGA
			CGTGGTGTTTGTCAGCAAAGATGTGGCCA
			AGCACTTGGGGTTCCAGTCAGCAGGGGAA
			GCCCTGAGGGGCTTGTATGGTCGTGTGAG
			GAAAGGGGCTGTGCTTGTCTGTGCCTGGG
			CTGAGGAGGGCGCCGACGCCCTGGGCCC
			TGATGGCAAACTGATCCACTCGGATGCTTT
			CCCGCCACCCCGCGTGGTGGATACCCTG
			GGGGCTGGAGACACCTTCAATGCCTCCGT
			CATCTTCAGCCTCTCCCAGGGGAGGAGCG
			TGCAGGAAGCACTGAGATTCGGATGCCAG
			GTGGCCGGCAAGAAGTGTGGCCAGCAGG
			GCTTTGATGGCATCGTGTCAGAGCCGGTG
			CGGTAGGAGGTGCCGGCTCCCCGCACAC
			TATGGAGGCTGACATTGCGGCTGCATCGC
			CTTCTCCCCTCCATCCAGCCTGGCATCCA
			GGTTGCCCTGCTCAGGGGACAGATGCAG
			GCTGTGGGGAGGACTCCGCCTGTGTCCT
			GTGTTCCCCACACGTCTCTCCCTGCAGAG
			CCTCAGAGCGAATAAATCTTCCTCGGAGC
			CAGCTTCCCCTGGCAGCTTCTGTCCTCGA
			TGTCTGAACTGCTCTGGCTGGGCATTCCT
			GAGGCTCTGACTCTCCAGTCCTCCCTCCT
			CGTGTGCATTCCCCAAATTAACCTCTCCAC
			CCAGGCCCAGAGGAGGGGCTGCCTGGGC
			TATAGCAGCAAGAAGTGCCCCAGGCTTGC
			CGCCAGCTCTGCCCTGGCTGGGGAGGAC
			ACTCAGTGCCCCATACCCAGCGAACCTGC
			CAAAGAACCAGAAGCCATGAGAGCTCTTT
			GGGGCCCTGCGTTGTGCAGACTCTATTCC
			CATAGCTCAGAAGCTGGGAGTCCACACGG
			CTGAGCCAAACTGACAGGCCAGTGGGGG
			GCGAGGGGGTGGGGCGCCTCCTCTGCCC
			TGCCCACCAGCCTGTGATTTGGTGGCGTC
			TTTGTTGTCCAAAAATATCTCCTCCCGCTG
			ACTGCCCCAGAGCCTGAAAGTCTCACCCG
			TGGAGCCCACCTTGGAATTAAGGGGATGC
			CTCAGCCACAAATGTGACCCAAGATAGAG
			TGTTGTCCTCAGGGAGGTCGGATCTGGAA
			CACATATTGGAATTGGGGCCAACTCCAATA
			TAGAGTGGATAAGGCCTTATAATGTAAAGA
			GCACATAAGGTAAAGGGCTTTAGAGTCAG
			ACAGACCTAGATTCAAATCTGCCATTTAAT
			TAGCTGCATGTCACCTGAGGGTACAGCGC
			TTAACACAATCCGCCTCAATTTCTTCATCT
			GTCAAATGGAGCCAATTCTGCCTGGCTAC
			AGAATTATTGCGAGGATAAAATCATGTA
Mouse	Mouse	N/A	GAGGGAGAGAACGCTTGCTTCTGTGCTCC	3
(Mm) KHK	(Mm)		GCCTGCGAAGGCGAAGTTTCTGTTGCCAG
nucleotide			ACTGTGCTAGTCCGGGTGGTCCAGGGTCT
sequence			GCAGCAGGCGCAGAGGGATCGGAAAGGC
NM_00843			GATGCATTACTAGTGCGCTTTCGCTTTGAC
9.4			AGCTGAGGCGGAAAAGTGAGAGGGCCTG
			CCATTGGCCGGGCTAGGTAACCCACCCTT
			GCAAAGCAGAAAGCTCCCTGCGGGAGGA
			GTTCTGCACGCAGAGGAGGAGCCAAGGTA
			GCCAGTGAGAAGTTGGGACACGGTCCTCC
			AGTAGATAAGAGGCAGAGCCCAGCAGGAA
			CCCCCTCTGCTTGCGGGTAGGAAGCTTGG
			GGAGCAGCCTCATGGAAGAGAAGCAGATC
			CTGTGCGTGGGGCTGGTGGTGCTGGACA
			TCATCAATGTGGTGGACAAATACCCAGAG
			GAAGACACGGATCGCAGGTGCCTGTCCCA
			GAGATGGCAGCGTGGAGGCAACGCATCC
			AACTCCTGCACTGTCCTTTCCTTGCTTGGA
			GCCCGCTGTGCCTTCATGGGCTCTTTGGC
			CCCTGGCCACGTTGCCGACTTCCTGGTGG
			CTGACTTCAGGCAGAGGGGCGTGGATGT
			GTCTCAAGTGACTTGGCAGAGCCAGGGAG
			ATACCCCTTGCTCTTGCTGCATCGTCAACA
			ACTCCAATGGCTCCCGTACCATTATACTCT
			ACGACACGAACCTGCCAGATGTGTCTGCT
			AAGGACTTTGAGAAGGTCGATCTGACCCG
			GTTCAAGTGGATCCACATTGAGGGCCGGA
			ATGCATCGGAACAGGTGAAGATGCTGCAG
			CGGATAGAGGAGCACAATGCCAAGCAGCC
			TCTGCCACAGAAGGTCCGGGTGTCGGTG
			GAGATAGAGAAGCCCCGTGAGGAGCTCTT
			CCAGTTGTTTAGCTATGGTGAGGTGGTGT
			TTGTCAGCAAAGATGTGGCCAAGCACCTG
			GGGTTCCAGTCAGCAGTGGAGGCCCTGA
			GGGGCTTGTACAGTCGAGTGAAGAAAGGG
			GCTACGCTTGTCTGTGCCTGGGCTGAGGA
			GGGTGCCGATGCCCTGGGCCCCGATGGT
			CAGCTGCTCCACTCAGATGCCTTCCCACC
			GCCCCGAGTAGTAGACACTCTTGGGGCTG
			GAGACACCTTCAATGCCTCTGTCATCTTCA
			GCCTCTCGAAGGGAAACAGCATGCAAGAG
			GCCCTGAGATTCGGGTGCCAGGTGGCTG
			GCAAGAAGTGTGGCTTGCAGGGGTTTGAT
			GGCATTGTGTGAGAGGCAAGCGGCACCA
			GCTCGATACCTCAGAGGCTGGCACCATGC
			CTGCCACTGCCTTCTCTACTTCCTCCAGCT
			TAGCATCCAGCTGCCATTCCCCGGCAGGT
			GTGGGATGTGGGACAGCCTCTGTCTGTGT
			CTGCGTCTCTGTATACCTATCTCCTCTCTG
			CAGATACCTGGAGCAAATAAATCTTCCCCT
			GAGCCAGC
KHK-115-	Hs-Mf	25 mer	AGCGCAUUUUCUCUUUGCAUUCUCG	4
154	commons	Sense
		Strand
KHK-116-	Hs-Mf	25 mer	GCGCAUUUUCUCUUUGCAUUCUCGA	5
155	commons	Sense
		Strand
KHK-117-	Hs-Mf	25 mer	CGCAUUUUCUCUUUGCAUUCUCGAG	6
156	commons	Sense
		Strand
KHK-118-	Hs-Mf	25 mer	GCAUUUUCUCUUUGCAUUCUCGAGA	7
157	commons	Sense
		Strand
KHK-119-	Hs-Mf	25 mer	CAUUUUCUCUUUGCAUUCUCGAGAT	8
158	commons	Sense
		Strand
KHK-120-	Hs-Mf	25 mer	AUUUUCUCUUUGCAUUCUCGAGATC	9
159	commons	Sense
		Strand
KHK-121-	Hs-Mf	25 mer	UUUUCUCUUUGCAUUCUCGAGAUCG	10
160	commons	Sense
		Strand
KHK-122-	Hs-Mf	25 mer	UUUCUCUUUGCAUUCUCGAGAUCGC	11
161	commons	Sense
		Strand
KHK-123-	Hs-Mf	25 mer	UUCUCUUUGCAUUCUCGAGAUCGCT	12
162	commons	Sense
		Strand
KHK-124-	Hs-Mf	25 mer	UCUCUUUGCAUUCUCGAGAUCGCTT	13
163	commons	Sense
		Strand
KHK-125-	Hs-Mf	25 mer	CUCUUUGCAUUCUCGAGAUCGCUTA	14
164	commons	Sense
		Strand
KHK-126-	Hs-Mf	25 mer	UCUUUGCAUUCUCGAGAUCGCUUAG	15
165	commons	Sense
		Strand
KHK-127-	Hs-Mf	25 mer	CUUUGCAUUCUCGAGAUCGCUUAGC	16
166	commons	Sense
		Strand
KHK-128-	Hs-Mf	25 mer	UUUGCAUUCUCGAGAUCGCUUAGCC	17
167	commons	Sense
		Strand
KHK-179	Hs	25 mer	GUGAGUCCAUCUGACAAGCGAGGAA	18
	unique	Sense
		Strand
KHK-181-	Hs-Mf	25 mer	GAGUCCAUCUGACAAGCGAGGAAAC	19
220	commons	Sense
		Strand
KHK-182-	Hs-Mf	25 mer	AGUCCAUCUGACAAGCGAGGAAACT	20
221	commons	Sense
		Strand
KHK-183-	Hs-Mf	25 mer	GUCCAUCUGACAAGCGAGGAAACTA	21
222	commons	Sense
		Strand
KHK-184-	Hs-Mf	25 mer	UCCAUCUGACAAGCGAGGAAACUAA	22
223	commons	Sense
		Strand
KHK-185-	Hs-Mf	25 mer	CCAUCUGACAAGCGAGGAAACUAAG	23
224	commons	Sense
		Strand
KHK-186-	Hs-Mf	25 mer	CAUCUGACAAGCGAGGAAACUAAGG	24
225	commons	Sense
		Strand
KHK-187-	Hs-Mf	25 mer	AUCUGACAAGCGAGGAAACUAAGGC	25
226	commons	Sense
		Strand
KHK-188-	Hs-Mf	25 mer	UCUGACAAGCGAGGAAACUAAGGCT	26
227	commons	Sense
		Strand
KHK-431-	Hs-Mf	25 mer	GGACACCAUCCUCCUGGAUAAGAGG	27
470	commons	Sense
		Strand
KHK-432-	Hs-Mf	25 mer	GACACCAUCCUCCUGGAUAAGAGGC	28
471	commons	Sense
		Strand
KHK-433-	Hs-Mf	25 mer	ACACCAUCCUCCUGGAUAAGAGGCA	29
472	commons	Sense
		Strand
KHK-507-	Hs-Mf-	25 mer	AGCCUCAUGGAAGAGAAGCAGAUCC	30
545-376-	Mm-Rn	Sense
218	commons	Strand
KHK-508-	Hs-Mf-	25 mer	GCCUCAUGGAAGAGAAGCAGAUCCT	31
546-377-	Mm-Rn	Sense
219	commons	Strand
KHK-509-	Hs-Mf-	25 mer	CCUCAUGGAAGAGAAGCAGAUCCTG	32
547-378-	Mm-Rn	Sense
220	commons	Strand
KHK-510-	Hs-Mf-	25 mer	CUCAUGGAAGAGAAGCAGAUCCUGT	33
548-379-	Mm-Rn	Sense
221	commons	Strand
KHK-511-	Hs-Mf-	25 mer	UCAUGGAAGAGAAGCAGAUCCUGTG	34
549-380-	Mm-Rn	Sense
222	commons	Strand
KHK-512-	Hs-Mf-	25 mer	CAUGGAAGAGAAGCAGAUCCUGUGC	35
550-381-	Mm-Rn	Sense
223	commons	Strand
KHK-513-	Hs-Mf-	25 mer	AUGGAAGAGAAGCAGAUCCUGUGCG	36
551-382-	Mm-Rn	Sense
224	commons	Strand
KHK-514-	Hs-Mf-	25 mer	UGGAAGAGAAGCAGAUCCUGUGCGT	37
552-383-	Mm-Rn	Sense
225	commons	Strand
KHK-515-	Hs-Mf-	25 mer	GGAAGAGAAGCAGAUCCUGUGCGTG	38
553-384-	Mm-Rn	Sense
226	commons	Strand
KHK-516-	Hs-Mf-	25 mer	GAAGAGAAGCAGAUCCUGUGCGUGG	39
554-385-	Mm-Rn	Sense
227	commons	Strand
KHK-517-	Hs-Mf-	25 mer	AAGAGAAGCAGAUCCUGUGCGUGGG	40
555-386-	Mm-Rn	Sense
228	commons	Strand
KHK-518-	Hs-Mf-	25 mer	AGAGAAGCAGAUCCUGUGCGUGGGG	41
556-387-	Mm-Rn	Sense
229	commons	Strand
KHK-520-	Hs-Mf-	25 mer	AGAAGCAGAUCCUGUGCGUGGGGCT	42
558-389-	Mm-Rn	Sense
231	commons	Strand
KHK-521-	Hs-Mf-	25 mer	GAAGCAGAUCCUGUGCGUGGGGCTA	43
559-390-	Mm-Rn	Sense
232	commons	Strand
KHK-522-	Hs-Mf-	25 mer	AAGCAGAUCCUGUGCGUGGGGCUAG	44
560-391-	Mm-Rn	Sense
233	commons	Strand
KHK-541-	Hs-Mf	25 mer	GGCUAGUGGUGCUGGACGUCAUCAG	45
579	commons	Sense
		Strand
KHK-544-	Hs-Mf	25 mer	UAGUGGUGCUGGACGUCAUCAGCCT	46
582	commons	Sense
		Strand
KHK-546-	Hs-Mf	25 mer	GUGGUGCUGGACGUCAUCAGCCUGG	47
584	commons	Sense
		Strand
KHK-547-	Hs-Mf	25 mer	UGGUGCUGGACGUCAUCAGCCUGGT	48
585	commons	Sense
		Strand
KHK-548-	Hs-Mf	25 mer	GGUGCUGGACGUCAUCAGCCUGGTG	49
586	commons	Sense
		Strand
KHK-549-	Hs-Mf	25 mer	GUGCUGGACGUCAUCAGCCUGGUGG	50
587	commons	Sense
		Strand
KHK-550-	Hs-Mf	25 mer	UGCUGGACGUCAUCAGCCUGGUGGA	51
588	commons	Sense
		Strand
KHK-551-	Hs-Mf	25 mer	GCUGGACGUCAUCAGCCUGGUGGAC	52
589	commons	Sense
		Strand
KHK-552-	Hs-Mf	25 mer	CUGGACGUCAUCAGCCUGGUGGACA	53
590	commons	Sense
		Strand
KHK-553-	Hs-Mf	25 mer	UGGACGUCAUCAGCCUGGUGGACAA	54
591	commons	Sense
		Strand
KHK-554-	Hs-Mf	25 mer	GGACGUCAUCAGCCUGGUGGACAAG	55
592	commons	Sense
		Strand
KHK-555-	Hs-Mf	25 mer	GACGUCAUCAGCCUGGUGGACAAGT	56
593	commons	Sense
		Strand
KHK-556-	Hs-Mf	25 mer	ACGUCAUCAGCCUGGUGGACAAGTA	57
594	commons	Sense
		Strand
KHK-557-	Hs-Mf	25 mer	CGUCAUCAGCCUGGUGGACAAGUAC	58
595	commons	Sense
		Strand
KHK-558-	Hs-Mf	25 mer	GUCAUCAGCCUGGUGGACAAGUACC	59
596	commons	Sense
		Strand
KHK-559-	Hs-Mf	25 mer	UCAUCAGCCUGGUGGACAAGUACCC	60
597	commons	Sense
		Strand
KHK-560-	Hs-Mf	25 mer	CAUCAGCCUGGUGGACAAGUACCCT	61
598	commons	Sense
		Strand
KHK-561-	Hs-Mf	25 mer	AUCAGCCUGGUGGACAAGUACCCTA	62
599	commons	Sense
		Strand
KHK-562-	Hs-Mf	25 mer	UCAGCCUGGUGGACAAGUACCCUAA	63
600	commons	Sense
		Strand
KHK-563-	Hs-Mf	25 mer	CAGCCUGGUGGACAAGUACCCUAAG	64
601	commons	Sense
		Strand
KHK-564-	Hs-Mf	25 mer	AGCCUGGUGGACAAGUACCCUAAGG	65
602	commons	Sense
		Strand
KHK-565-	Hs-Mf	25 mer	GCCUGGUGGACAAGUACCCUAAGGA	66
603	commons	Sense
		Strand
KHK-566-	Hs-Mf	25 mer	CCUGGUGGACAAGUACCCUAAGGAG	67
604	commons	Sense
		Strand
KHK-567-	Hs-Mf	25 mer	CUGGUGGACAAGUACCCUAAGGAGG	68
605	commons	Sense
		Strand
KHK-568-	Hs-Mf	25 mer	UGGUGGACAAGUACCCUAAGGAGGA	69
606	commons	Sense
		Strand
KHK-569-	Hs-Mf	25 mer	GGUGGACAAGUACCCUAAGGAGGAC	70
607	commons	Sense
		Strand
KHK-570-	Hs-Mf	25 mer	GUGGACAAGUACCCUAAGGAGGACT	71
608	commons	Sense
		Strand
KHK-571-	Hs-Mf	25 mer	UGGACAAGUACCCUAAGGAGGACTC	72
609	commons	Sense
		Strand
KHK-572-	Hs-Mf	25 mer	GGACAAGUACCCUAAGGAGGACUCG	73
610	commons	Sense
		Strand
KHK-573-	Hs-Mf	25 mer	GACAAGUACCCUAAGGAGGACUCGG	74
611	commons	Sense
		Strand
KHK-574-	Hs-Mf	25 mer	ACAAGUACCCUAAGGAGGACUCGGA	75
612	commons	Sense
		Strand
KHK-575-	Hs-Mf	25 mer	CAAGUACCCUAAGGAGGACUCGGAG	76
613	commons	Sense
		Strand
KHK-576-	Hs-Mf	25 mer	AAGUACCCUAAGGAGGACUCGGAGA	77
614	commons	Sense
		Strand
KHK-577-	Hs-Mf	25 mer	AGUACCCUAAGGAGGACUCGGAGAT	78
615	commons	Sense
		Strand
KHK-638-	Hs-Mf	25 mer	CGCGUCCAACUCCUGCACCGUUCTC	79
676	commons	Sense
		Strand
KHK-641-	Hs-Mf	25 mer	GUCCAACUCCUGCACCGUUCUCUCC	80
679	commons	Sense
		Strand
KHK-642-	Hs-Mf	25 mer	UCCAACUCCUGCACCGUUCUCUCCC	81
680	commons	Sense
		Strand
KHK-643-	Hs-Mf	25 mer	CCAACUCCUGCACCGUUCUCUCCCT	82
681	commons	Sense
		Strand
KHK-644-	Hs-Mf	25 mer	CAACUCCUGCACCGUUCUCUCCCTG	83
682	commons	Sense
		Strand
KHK-645-	Hs-Mf	25 mer	AACUCCUGCACCGUUCUCUCCCUGC	84
683	commons	Sense
		Strand
KHK-646-	Hs-Mf	25 mer	ACUCCUGCACCGUUCUCUCCCUGCT	85
684	commons	Sense
		Strand
KHK-647-	Hs-Mf	25 mer	CUCCUGCACCGUUCUCUCCCUGCTC	86
685	commons	Sense
		Strand
KHK-650-	Hs-Mf	25 mer	CUGCACCGUUCUCUCCCUGCUCGGA	87
688	commons	Sense
		Strand
KHK-676-	Hs-Mf	25 mer	CCCCCUGUGCCUUCAUGGGCUCAAT	88
714	commons	Sense
		Strand
KHK-713-	Hs-Mf	25 mer	UGUUGCUGACUUCCUGGUGGCCGAC	89
722	commons	Sense
		Strand
KHK-826-	Hs-Mf	25 mer	AUGGCAACCGUACCAUUGUGCUCCA	90
835	commons	Sense
		Strand
KHK-827-	Hs-Mf	25 mer	UGGCAACCGUACCAUUGUGCUCCAT	91
836	commons	Sense
		Strand
KHK-829-	Hs-Mf	25 mer	GCAACCGUACCAUUGUGCUCCAUGA	92
838	commons	Sense
		Strand
KHK-830-	Hs-Mf	25 mer	CAACCGUACCAUUGUGCUCCAUGAC	93
839	commons	Sense
		Strand
KHK-831-	Hs-Mf	25 mer	AACCGUACCAUUGUGCUCCAUGACA	94
840	commons	Sense
		Strand
KHK-832-	Hs-Mf	25 mer	ACCGUACCAUUGUGCUCCAUGACAC	95
841	commons	Sense
		Strand
KHK-857-	Hs-Mf	25 mer	GAGCCUGCCAGAUGUGUCUGCUACA	96
895	commons	Sense
		Strand
KHK-858-	Hs-Mf	25 mer	AGCCUGCCAGAUGUGUCUGCUACAG	97
896	commons	Sense
		Strand
KHK-859-	Hs-Mf	25 mer	GCCUGCCAGAUGUGUCUGCUACAGA	98
897	commons	Sense
		Strand
KHK-860-	Hs-Mf-	25 mer	CCUGCCAGAUGUGUCUGCUACAGAC	99
898-729-	Mm-Rn	Sense
571	commons	Strand
KHK-861-	Hs-Mf-	25 mer	CUGCCAGAUGUGUCUGCUACAGACT	100
899-730-	Mm-Rn	Sense
572	commons	Strand
KHK-862-	Hs-Mf	25 mer	UGCCAGAUGUGUCUGCUACAGACTT	101
900	commons	Sense
		Strand
KHK-865	Hs	25 mer	CAGAUGUGUCUGCUACAGACUUUGA	102
	unique	Sense
		Strand
KHK-880	Hs	25 mer	CAGACUUUGAGAAGGUUGAUCUGAC	103
	unique	Sense
		Strand
KHK-882-	Hs-Mf	25 mer	GACUUUGAGAAGGUUGAUCUGACCC	104
920	commons	Sense
		Strand
KHK-883-	Hs-Mf	25 mer	ACUUUGAGAAGGUUGAUCUGACCCA	105
921	commons	Sense
		Strand
KHK-884-	Hs-Mf	25 mer	CUUUGAGAAGGUUGAUCUGACCCAG	106
922	commons	Sense
		Strand
KHK-885-	Hs-Mf	25 mer	UUUGAGAAGGUUGAUCUGACCCAGT	107
923	commons	Sense
		Strand
KHK-886-	Hs-Mf	25 mer	UUGAGAAGGUUGAUCUGACCCAGTT	108
924	commons	Sense
		Strand
KHK-887-	Hs-Mf	25 mer	UGAGAAGGUUGAUCUGACCCAGUTC	109
925	commons	Sense
		Strand
KHK-888-	Hs-Mf	25 mer	GAGAAGGUUGAUCUGACCCAGUUCA	110
926	commons	Sense
		Strand
KHK-889-	Hs-Mf	25 mer	AGAAGGUUGAUCUGACCCAGUUCAA	111
927	commons	Sense
		Strand
KHK-890-	Hs-Mf	25 mer	GAAGGUUGAUCUGACCCAGUUCAAG	112
928	commons	Sense
		Strand
KHK-891-	Hs-Mf	25 mer	AAGGUUGAUCUGACCCAGUUCAAGT	113
929	commons	Sense
		Strand
KHK-892-	Hs-Mf	25 mer	AGGUUGAUCUGACCCAGUUCAAGTG	114
930	commons	Sense
		Strand
KHK-893-	Hs-Mf	25 mer	GGUUGAUCUGACCCAGUUCAAGUGG	115
931	commons	Sense
		Strand
KHK-894-	Hs-Mf	25 mer	GUUGAUCUGACCCAGUUCAAGUGGA	116
932	commons	Sense
		Strand
KHK-895-	Hs-Mf	25 mer	UUGAUCUGACCCAGUUCAAGUGGAT	117
933	commons	Sense
		Strand
KHK-896-	Hs-Mf	25 mer	UGAUCUGACCCAGUUCAAGUGGATC	118
934	commons	Sense
		Strand
KHK-897-	Hs-Mf	25 mer	GAUCUGACCCAGUUCAAGUGGAUCC	119
935	commons	Sense
		Strand
KHK-898-	Hs-Mf	25 mer	AUCUGACCCAGUUCAAGUGGAUCCA	120
936	commons	Sense
		Strand
KHK-899-	Hs-Mf	25 mer	UCUGACCCAGUUCAAGUGGAUCCAC	121
937	commons	Sense
		Strand
KHK-900-	Hs-Mf	25 mer	CUGACCCAGUUCAAGUGGAUCCACA	122
938	commons	Sense
		Strand
KHK-901-	Hs-Mf	25 mer	UGACCCAGUUCAAGUGGAUCCACAT	123
939	commons	Sense
		Strand
KHK-902-	Hs-Mf	25 mer	GACCCAGUUCAAGUGGAUCCACATT	124
940	commons	Sense
		Strand
KHK-903-	Hs-Mf	25 mer	ACCCAGUUCAAGUGGAUCCACAUTG	125
941	commons	Sense
		Strand
KHK-904-	Hs-Mf	25 mer	CCCAGUUCAAGUGGAUCCACAUUGA	126
942	commons	Sense
		Strand
KHK-905-	Hs-Mf	25 mer	CCAGUUCAAGUGGAUCCACAUUGAG	127
943	commons	Sense
		Strand
KHK-906-	Hs-Mf	25 mer	CAGUUCAAGUGGAUCCACAUUGAGG	128
944	commons	Sense
		Strand
KHK-907-	Hs-Mf	25 mer	AGUUCAAGUGGAUCCACAUUGAGGG	129
945	commons	Sense
		Strand
KHK-908-	Hs-Mf-	25 mer	GUUCAAGUGGAUCCACAUUGAGGGC	130
946-777-	Mm-Rn	Sense
619	commons	Strand
KHK-909-	Hs-Mf-	25 mer	UUCAAGUGGAUCCACAUUGAGGGCC	131
947-778-	Mm-Rn	Sense
620	commons	Strand
KHK-910-	Hs-Mf-	25 mer	UCAAGUGGAUCCACAUUGAGGGCCG	132
948-779-	Mm-Rn	Sense
621	commons	Strand
KHK-911-	Hs-Mf-	25 mer	CAAGUGGAUCCACAUUGAGGGCCGG	133
949-780-	Mm-Rn	Sense
622	commons	Strand
KHK-912-	Hs-Mf-	25 mer	AAGUGGAUCCAGAUUGAGGGCCGGA	134
950-781-	Mm-Rn	Sense
623	commons	Strand
KHK-913-	Hs-Mf-	25 mer	AGUGGAUCCACAUUGAGGGCCGGAA	135
951-782-	Mm-Rn	Sense
624	commons	Strand
KHK-914-	Hs-Mf-	25 mer	GUGGAUCCACAUUGAGGGCCGGAAC	136
952-783-	Mm-Rn	Sense
625	commons	Strand
KHK-939-	Hs-Mf	25 mer	GCAUCGGAGCAGGUGAAGAUGCUGC	137
977	commons	Sense
		Strand
KHK-940-	Hs-Mf	25 mer	CAUCGGAGCAGGUGAAGAUGCUGCA	138
978	commons	Sense
		Strand
KHK-941-	Hs-Mf	25 mer	AUCGGAGCAGGUGAAGAUGCUGCAG	139
979	commons	Sense
		Strand
KHK-942-	Hs-Mf	25 mer	UCGGAGCAGGUGAAGAUGCUGCAGC	140
980	commons	Sense
		Strand
KHK-943-	Hs-Mf	25 mer	CGGAGCAGGUGAAGAUGCUGCAGCG	141
981	commons	Sense
		Strand
KHK-944-	Hs-Mf	25 mer	GGAGCAGGUGAAGAUGCUGCAGCGG	142
982	commons	Sense
		Strand
KHK-945-	Hs-Mf	25 mer	GAGCAGGUGAAGAUGCUGCAGCGGA	143
983	commons	Sense
		Strand
KHK-946-	Hs-Mf	25 mer	AGCAGGUGAAGAUGCUGCAGCGGAT	144
984	commons	Sense
		Strand
KHK-947-	Hs-Mf	25 mer	GCAGGUGAAGAUGCUGCAGCGGATA	145
985	commons	Sense
		Strand
KHK-948-	Hs-Mf-	25 mer	CAGGUGAAGAUGCUGCAGCGGAUAG	146
986-817	Mm	Sense
	commons	Strand
KHK-949-	Hs-Mf-	25 mer	AGGUGAAGAUGCUGCAGCGGAUAGA	147
987-818	Mm	Sense
	commons	Strand
KHK-950-	Hs-Mf-	25 mer	GGUGAAGAUGCUGCAGCGGAUAGAC	148
988-819	Mm	Sense
	commons	Strand
KHK-951-	Hs-Mf-	25 mer	GUGAAGAUGCUGCAGCGGAUAGACG	149
989-820	Mm	Sense
	commons	Strand
KHK-952-	Hs-Mf-	25 mer	UGAAGAUGCUGCAGCGGAUAGACGC	150
990-821	Mm	Sense
	commons	Strand
KHK-953-	Hs-Mf-	25 mer	GAAGAUGCUGCAGCGGAUAGACGCA	151
991-822	Mm	Sense
	commons	Strand
KHK-954-	Hs-Mf-	25 mer	AAGAUGCUGCAGCGGAUAGACGCAC	152
992-823	Mm	Sense
	commons	Strand
KHK-955-	Hs-Mf-	25 mer	AGAUGCUGCAGCGGAUAGACGCACA	153
993-824	Mm	Sense
	commons	Strand
KHK-956-	Hs-Mf	25 mer	GAUGCUGCAGCGGAUAGACGCACAC	154
994	commons	Sense
		Strand
KHK-957-	Hs-Mf	25 mer	AUGCUGCAGCGGAUAGACGCACACA	155
995	commons	Sense
		Strand
KHK-958-	Hs-Mf	25 mer	UGCUGCAGCGGAUAGACGCACACAA	156
996	commons	Sense
		Strand
KHK-978-	Hs-Mf	25 mer	CACAACACCAGGCAGCCUCCAGAGC	157
1016	commons	Sense
		Strand
KHK-982-	Hs-Mf	25 mer	ACACCAGGCAGCCUCCAGAGCAGAA	158
1020	commons	Sense
		Strand
KHK-983-	Hs-Mf	25 mer	CACCAGGCAGCCUCCAGAGCAGAAG	159
1021	commons	Sense
		Strand
KHK-984-	Hs-Mf	25 mer	ACCAGGCAGCCUCCAGAGCAGAAGA	160
1022	commons	Sense
		Strand
KHK-985-	Hs-Mf	25 mer	CCAGGCAGCCUCCAGAGCAGAAGAT	161
1023	commons	Sense
		Strand
KHK-991-	Hs-Mf	25 mer	AGCCUCCAGAGCAGAAGAUCCGGGT	162
1029	commons	Sense
		Strand
KHK-992-	Hs-Mf	25 mer	GCCUCCAGAGCAGAAGAUCCGGGTG	163
1030	commons	Sense
		Strand
KHK-993-	Hs-Mf	25 mer	CCUCCAGAGCAGAAGAUCCGGGUGT	164
1031	commons	Sense
		Strand
KHK-999-	Hs-Mf	25 mer	GAGCAGAAGAUCCGGGUGUCCGUGG	165
1037	commons	Sense
		Strand
KHK-1000-	Hs-Mf	25 mer	AGCAGAAGAUCCGGGUGUCCGUGGA	166
1038	commons	Sense
		Strand
KHK-1019-	Hs-Mf	25 mer	CGUGGAGGUGGAGAAGCCACGAGAG	167
1057	commons	Sense
		Strand
KHK-1054-	Hs-Mf	25 mer	AGCUGUUUGGCUACGGAGACGUGGT	168
1092	commons	Sense
		Strand
KHK-1055-	Hs-Mf	25 mer	GCUGUUUGGCUACGGAGACGUGGTG	169
1093	commons	Sense
		Strand
KHK-1057-	Hs-Mf	25 mer	UGUUUGGCUACGGAGACGUGGUGTT	170
1095	commons	Sense
		Strand
KHK-1058-	Hs-Mf	25 mer	GUUUGGCUACGGAGACGUGGUGUTT	171
1096	commons	Sense
		Strand
KHK-1059-	Hs-Mf	25 mer	UUUGGCUACGGAGACGUGGUGUUTG	172
1097	commons	Sense
		Strand
KHK-1060-	Hs-Mf	25 mer	UUGGCUACGGAGACGUGGUGUUUGT	173
1098	commons	Sense
		Strand
KHK-1061-	Hs-Mf	25 mer	UGGCUACGGAGACGUGGUGUUUGTC	174
1099	commons	Sense
		Strand
KHK-1062-	Hs-Mf	25 mer	GGCUACGGAGACGUGGUGUUUGUCA	175
1100	commons	Sense
		Strand
KHK-1063-	Hs-Mf	25 mer	GCUACGGAGACGUGGUGUUUGUCAG	176
1101	commons	Sense
		Strand
KHK-1064-	Hs-Mf	25 mer	CUACGGAGACGUGGUGUUUGUCAGC	177
1102	commons	Sense
		Strand
KHK-1065-	Hs-Mf	25 mer	UACGGAGACGUGGUGUUUGUCAGCA	178
1103	commons	Sense
		Strand
KHK-1066-	Hs-Mf	25 mer	ACGGAGACGUGGUGUUUGUCAGCAA	179
1104	commons	Sense
		Strand
KHK-1067-	Hs-Mf	25 mer	CGGAGACGUGGUGUUUGUCAGCAAA	180
1105	commons	Sense
		Strand
KHK-1068-	Hs-Mf	25 mer	GGAGACGUGGUGUUUGUCAGCAAAG	181
1106	commons	Sense
		Strand
KHK-1069-	Hs-Mf	25 mer	GAGACGUGGUGUUUGUCAGCAAAGA	182
1107	commons	Sense
		Strand
KHK-1070-	Hs-Mf	25 mer	AGACGUGGUGUUUGUCAGCAAAGAT	183
1108	commons	Sense
		Strand
KHK-1071-	Hs-Mf	25 mer	GACGUGGUGUUUGUCAGCAAAGATG	184
1109	commons	Sense
		Strand
KHK-1072-	Hs-Mf	25 mer	ACGUGGUGUUUGUCAGCAAAGAUGT	185
1110	commons	Sense
		Strand
KHK-1073-	Hs-Mf	25 mer	CGUGGUGUUUGUCAGCAAAGAUGTG	186
1111	commons	Sense
		Strand
KHK-1074-	Hs-Mf-	25 mer	GUGGUGUUUGUCAGCAAAGAUGUGG	187
1112-943-	Mm-Rn	Sense
785	commons	Strand
KHK-1075-	Hs-Mf-	25 mer	UGGUGUUUGUCAGCAAAGAUGUGGC	188
1113-944-	Mm-Rn	Sense
786	commons	Strand
KHK-1076-	Hs-Mf-	25 mer	GGUGUUUGUCAGCAAAGAUGUGGCC	189
1114-945-	Mm-Rn	Sense
787	commons	Strand
KHK-1077-	Hs-Mf-	25 mer	GUGUUUGUCAGCAAAGAUGUGGCCA	190
1115-946-	Mm-Rn	Sense
788	commons	Strand
KHK-1078-	Hs-Mf-	25 mer	UGUUUGUCAGCAAAGAUGUGGCCAA	191
1116-947-	Mm-Rn	Sense
789	commons	Strand
KHK-1079-	Hs-Mf-	25 mer	GUUUGUCAGCAAAGAUGUGGCCAAG	192
1117-948-	Mm-Rn	Sense
790	commons	Strand
KHK-1080-	Hs-Mf-	25 mer	UUUGUCAGCAAAGAUGUGGCCAAGC	193
11 18-949-	Mm-Rn	Sense
791	commons	Strand
KHK-1081-	Hs-Mf-	25 mer	UUGUCAGCAAAGAUGUGGCCAAGCA	194
1119-950-	Mm-Rn	Sense
792	commons	Strand
KHK-1082-	Hs-Mf-	25 mer	UGUCAGCAAAGAUGUGGCCAAGCAC	195
1120-951-	Mm-Rn	Sense
793	commons	Strand
KHK-1083-	Hs-Mf-	25 mer	GUCAGCAAAGAUGUGGCCAAGCACT	196
1121-952-	Mm-Rn	Sense
794	commons	Strand
KHK-1084-	Hs-Mf-	25 mer	UCAGCAAAGAUGUGGCCAAGCACTT	197
1122-953-	Mm-Rn	Sense
795	commons	Strand
KHK-1085-	Hs-Mf-	25 mer	CAGCAAAGAUGUGGCCAAGCACUTG	198
1123-954-	Mm-Rn	Sense
796	commons	Strand
KHK-1086-	Hs-Mf-	25 mer	AGCAAAGAUGUGGCCAAGCACUUGG	199
1124-955-	Mm-Rn	Sense
797	commons	Strand
KHK-1087-	Hs-Mf-	25 mer	GCAAAGAUGUGGCCAAGCACUUGGG	200
1125-956-	Mm-Rn	Sense
798	commons	Strand
KHK-1090-	Hs-Mf	25 mer	AAGAUGUGGCCAAGCACUUGGGGTT	201
1128	commons	Sense
		Strand
KHK-1091-	Hs-Mf	25 mer	AGAUGUGGCCAAGCACUUGGGGUTC	202
1129	commons	Sense
		Strand
KHK-1092-	Hs-Mf	25 mer	GAUGUGGCCAAGCACUUGGGGUUCC	203
1130	commons	Sense
		Strand
KHK-1093-	Hs-Mf	25 mer	AUGUGGCCAAGCACUUGGGGUUCCA	204
1131	commons	Sense
		Strand
KHK-1095-	Hs-Mf	25 mer	GUGGCCAAGCACUUGGGGUUCCAGT	205
1133	commons	Sense
		Strand
KHK-1096-	Hs-Mf	25 mer	UGGCCAAGCACUUGGGGUUCCAGTC	206
1134	commons	Sense
		Strand
KHK-1097-	Hs-Mf	25 mer	GGCCAAGCACUUGGGGUUCCAGUCA	207
1135	commons	Sense
		Strand
KHK-1099-	Hs-Mf	25 mer	CCAAGCACUUGGGGUUCCAGUCAGC	208
1137	commons	Sense
		Strand
KHK-1100-	Hs-Mf	25 mer	CAAGCACUUGGGGUUCCAGUCAGCA	209
1138	commons	Sense
		Strand
KHK-1101-	Hs-Mf	25 mer	AAGCACUUGGGGUUCCAGUCAGCAG	210
1139	commons	Sense
		Strand
KHK-1102-	Hs-Mf	25 mer	AGCACUUGGGGUUCCAGUCAGCAGA	211
1140	commons	Sense
		Strand
KHK-1103-	Hs-Mf	25 mer	GCACUUGGGGUUCCAGUCAGCAGAG	212
1141	commons	Sense
		Strand
KHK-1104-	Hs-Mf	25 mer	CACUUGGGGUUCCAGUCAGCAGAGG	213
1142	commons	Sense
		Strand
KHK-1106-	Hs-Mf	25 mer	CUUGGGGUUCCAGUCAGCAGAGGAA	214
1144	commons	Sense
		Strand
KHK-1107-	Hs-Mf	25 mer	UUGGGGUUCCAGUCAGCAGAGGAAG	215
1145	commons	Sense
		Strand
KHK-1135-	Hs-Mf	25 mer	UGAGGGGCUUGUAUGGUCGUGUGAG	216
1173	commons	Sense
		Strand
KHK-1136-	Hs-Mf	25 mer	GAGGGGCUUGUAUGGUCGUGUGAGG	217
ll74	commons	Sense
		Strand
KHK-1137-	Hs-Mf	25 mer	AGGGGCUUGUAUGGUCGUGUGAGGA	218
1175	commons	Sense
		Strand
KHK-1138-	Hs-Mf	25 mer	GGGGCUUGUAUGGUCGUGUGAGGAA	219
1176	commons	Sense
		Strand
KHK-1139-	Hs-Mf	25 mer	GGGCUUGUAUGGUCGUGUGAGGAAA	220
1177	commons	Sense
		Strand
KHK-1140-	Hs-Mf	25 mer	GGCUUGUAUGGUCGUGUGAGGAAAG	221
1178	commons	Sense
		Strand
KHK-1141-	Hs-Mf	25 mer	GCUUGUAUGGUCGUGUGAGGAAAGG	222
1179	commons	Sense
		Strand
KHK-1142-	Hs-Mf	25 mer	CUUGUAUGGUCGUGUGAGGAAAGGG	223
1180	commons	Sense
		Strand
KHK-1143-	Hs-Mf	25 mer	UUGUAUGGUCGUGUGAGGAAAGGGG	224
1181	commons	Sense
		Strand
KHK-1144-	Hs-Mf	25 mer	UGUAUGGUCGUGUGAGGAAAGGGGC	225
1182	commons	Sense
		Strand
KHK-1145-	Hs-Mf	25 mer	GUAUGGUCGUGUGAGGAAAGGGGCT	226
1183	commons	Sense
		Strand
KHK-1146-	Hs-Mf	25 mer	UAUGGUCGUGUGAGGAAAGGGGCTG	227
1184	commons	Sense
		Strand
KHK-1147-	Hs-Mf	25 mer	AUGGUCGUGUGAGGAAAGGGGCUGT	228
1185	commons	Sense
		Strand
KHK-1148-	Hs-Mf	25 mer	UGGUCGUGUGAGGAAAGGGGCUGTG	229
1186	commons	Sense
		Strand
KHK-1149-	Hs-Mf	25 mer	GGUCGUGUGAGGAAAGGGGCUGUGC	230
1187	commons	Sense
		Strand
KHK-1153-	Hs-Mf	25 mer	GUGUGAGGAAAGGGGCUGUGCUUGT	231
1191	commons	Sense
		Strand
KHK-1154-	Hs-Mf	25 mer	UGUGAGGAAAGGGGCUGUGCUUGTC	232
1192	commons	Sense
		Strand
KHK-1157-	Hs-Mf	25 mer	GAGGAAAGGGGCUGUGCUUGUCUGT	233
1195	commons	Sense
		Strand
KHK-1158-	Hs-Mf	25 mer	AGGAAAGGGGCUGUGCUUGUCUGTG	234
1196	commons	Sense
		Strand
KHK-1159-	Hs-Mf	25 mer	GGAAAGGGGCUGUGCUUGUCUGUGC	235
1197	commons	Sense
		Strand
KHK-1161-	Hs-Mf	25 mer	AAAGGGGCUGUGCUUGUCUGUGCCT	236
1199	commons	Sense
		Strand
KHK-1163-	Hs-Mf	25 mer	AGGGGCUGUGCUUGUCUGUGCCUGG	237
1201	commons	Sense
		Strand
KHK-1164-	Hs-Mf	25 mer	GGGGCUGUGCUUGUCUGUGCCUGGG	238
1202	commons	Sense
		Strand
KHK-1232-	Hs-Mf	25 mer	CCACUCGGAUGCUUUCCCGCCACCC	239
1270	commons	Sense
		Strand
KHK-1278-	Hs-Mf-	25 mer	GCUGGAGACACCUUCAAUGCCUCCG	240
1316-1147-	Mm-Rn	Sense
989	commons	Strand
KHK-1279-	Hs-Mf-	25 mer	CUGGAGACACCUUCAAUGCCUCCGT	241
1317-1148-	Mm-Rn	Sense
990	commons	Strand
KHK-1280-	Hs-Mf-	25 mer	UGGAGACACCUUCAAUGCCUCCGTC	242
1318-1149-	Mm-Rn	Sense
991	commons	Strand
KHK-1281-	Hs-Mf-	25 mer	GGAGACACCUUCAAUGCCUCCGUCA	243
1319-1150-	Mm-Rn	Sense
992	commons	Strand
KHK-1282-	Hs-Mf-	25 mer	GAGACACCUUCAAUGCCUCCGUCAT	244
1320-1151-	Mm-Rn	Sense
993	commons	Strand
KHK-1283-	Hs-Mf	25 mer	AGACACCUUCAAUGCCUCCGUCATC	245
1321	commons	Sense
		Strand
KHK-1284-	Hs-Mf	25 mer	GACACCUUCAAUGCCUCCGUCAUCT	246
1322	commons	Sense
		Strand
KHK-1285-	Hs-Mf	25 mer	ACACCUUCAAUGCCUCCGUCAUCTT	247
1323	commons	Sense
		Strand
KHK-1286-	Hs-Mf	25 mer	CACCUUCAAUGCCUCCGUCAUCUTC	248
1324	commons	Sense
		Strand
KHK-1287-	Hs-Mf	25 mer	ACCUUCAAUGCCUCCGUCAUCUUCA	249
1325	commons	Sense
		Strand
KHK-1288-	Hs-Mf	25 mer	CCUUCAAUGCCUCCGUCAUCUUCAG	250
1326	commons	Sense
		Strand
KHK-1289-	Hs-Mf	25 mer	CUUCAAUGCCUCCGUCAUCUUCAGC	251
1327	commons	Sense
		Strand
KHK-1290-	Hs-Mf	25 mer	UUCAAUGCCUCCGUCAUCUUCAGCC	252
1328	commons	Sense
		Strand
KHK-1291-	Hs-Mf	25 mer	UCAAUGCCUCCGUCAUCUUCAGCCT	253
1329	commons	Sense
		Strand
KHK-1292-	Hs-Mf	25 mer	CAAUGCCUCCGUCAUCUUCAGCCTC	254
1330	commons	Sense
		Strand
KHK-1293-	Hs-Mf	25 mer	AAUGCCUCCGUCAUCUUCAGCCUCT	255
1331	commons	Sense
		Strand
KHK-1294-	Hs-Mf	25 mer	AUGCCUCCGUCAUCUUCAGCCUCTC	256
1332	commons	Sense
		Strand
KHK-1295-	Hs-Mf	25 mer	UGCCUCCGUCAUCUUCAGCCUCUCC	257
1333	commons	Sense
		Strand
KHK-1297-	Hs-Mf	25 mer	CCUCCGUCAUCUUCAGCCUCUCCCA	258
1335	commons	Sense
		Strand
KHK-1323-	Hs-Mf	25 mer	GGGAGGAGCGUGCAGGAAGCACUGA	259
1361	commons	Sense
		Strand
KHK-1325-	Hs-Mf	25 mer	GAGGAGCGUGCAGGAAGCACUGAGA	260
1363	commons	Sense
		Strand
KHK-1326-	Hs-Mf	25 mer	AGGAGCGUGCAGGAAGCACUGAGAT	261
1364	commons	Sense
		Strand
KHK-1327-	Hs-Mf	25 mer	GGAGCGUGCAGGAAGCACUGAGATT	262
1365	commons	Sense
		Strand
KHK-1328-	Hs-Mf	25 mer	GAGCGUGCAGGAAGCACUGAGAUTC	263
1366	commons	Sense
		Strand
KHK-1329-	Hs-Mf	25 mer	AGCGUGCAGGAAGCACUGAGAUUCG	264
1367	commons	Sense
		Strand
KHK-1330-	Hs-Mf	25 mer	GCGUGCAGGAAGCACUGAGAUUCGG	265
1368	commons	Sense
		Strand
KHK-1331-	Hs-Mf	25 mer	CGUGCAGGAAGCACUGAGAUUCGGG	266
1369	commons	Sense
		Strand
KHK-1332-	Hs-Mf	25 mer	GUGCAGGAAGCACUGAGAUUCGGGT	267
1370	commons	Sense
		Strand
KHK-1333-	Hs-Mf	25 mer	UGCAGGAAGCACUGAGAUUCGGGTG	268
1371	commons	Sense
		Strand
KHK-1334-	Hs-Mf	25 mer	GCAGGAAGCACUGAGAUUCGGGUGC	269
1372	commons	Sense
		Strand
KHK-1335-	Hs-Mf	25 mer	CAGGAAGCACUGAGAUUCGGGUGCC	270
1373	commons	Sense
		Strand
KHK-1336-	Hs-Mf	25 mer	AGGAAGCACUGAGAUUCGGGUGCCA	271
1374	commons	Sense
		Strand
KHK-1385-	Hs-Mf	25 mer	GCAGGGCUUUGAUGGCAUCGUGUGA	272
1423	commons	Sense
		Strand
KHK-1387-	Hs-Mf	25 mer	AGGGCUUUGAUGGCAUCGUGUGAGA	273
1425	commons	Sense
		Strand
KHK-1388-	Hs-Mf	25 mer	GGGCUUUGAUGGCAUCGUGUGAGAG	274
1426	commons	Sense
		Strand
KHK-1389-	Hs-Mf	25 mer	GGCUUUGAUGGCAUCGUGUGAGAGC	275
1427	commons	Sense
		Strand
KHK-1538-	Hs-Mf	25 mer	GCCUGUGUCCUGUGUUCCCCACAGG	276
1588	commons	Sense
		Strand
KHK-1540-	Hs-Mf	25 mer	CUGUGUCCUGUGUUCCCCACAGGGA	277
1590	commons	Sense
		Strand
KHK-1542-	Hs-Mf	25 mer	GUGUCCUGUGUUCCCCACAGGGAGA	278
1592	commons	Sense
		Strand
KHK-1665-	Hs-Mf	25 mer	CAUUCCUGAGGCUCUGACUCUUCGA	279
1708	commons	Sense
		Strand
KHK-1666-	Hs-Mf	25 mer	AUUCCUGAGGCUCUGACUCUUCGAT	280
1709	commons	Sense
		Strand
KHK-1667-	Hs-Mf	25 mer	UUCCUGAGGCUCUGACUCUUCGATC	281
1710	commons	Sense
		Strand
KHK-1707-	Hs-Mf	25 mer	CAUUCCCCAAAUUAACCUCUCCGCC	282
1750	commons	Sense
		Strand
KHK-1708-	Hs-Mf	25 mer	AUUCCCCAAAUUAACCUCUCCGCCC	283
1751	commons	Sense
		Strand
KHK-1709-	Hs-Mf	25 mer	UUCCCCAAAUUAACCUCUCCGCCCA	284
1752	commons	Sense
		Strand
KHK-1869-	Hs-Mf	25 mer	GGGCCCUGCGUUGUGCAGACUCUAT	285
1918	commons	Sense
		Strand
KHK-1870-	Hs-Mf	25 mer	GGCCCUGCGUUGUGCAGACUCUATT	286
1919	commons	Sense
		Strand
KHK-1871-	Hs-Mf	25 mer	GCCCUGCGUUGUGCAGACUCUAUTC	287
1920	commons	Sense
		Strand
KHK-1872-	Hs-Mf	25 mer	CCCUGCGUUGUGCAGACUCUAUUCC	288
1921	commons	Sense
		Strand
KHK-1873-	Hs-Mf	25 mer	CCUGCGUUGUGCAGACUCUAUUCCC	289
1922	commons	Sense
		Strand
KHK-1874-	Hs-Mf	25 mer	CUGCGUUGUGCAGACUCUAUUCCCA	290
1923	commons	Sense
		Strand
KHK-1875-	Hs-Mf	25 mer	UGCGUUGUGCAGACUCUAUUCCCAC	291
1924	commons	Sense
		Strand
KHK-1876-	Hs-Mf	25 mer	GCGUUGUGCAGACUCUAUUCCCACA	292
1925	commons	Sense
		Strand
KHK-1877-	Hs-Mf	25 mer	CGUUGUGCAGACUCUAUUCCCACAG	293
1926	commons	Sense
		Strand
KHK-1878-	Hs-Mf	25 mer	GUUGUGCAGACUCUAUUCCCACAGC	294
1927	commons	Sense
		Strand
KHK-1879-	Hs-Mf	25 mer	UUGUGCAGACUCUAUUCCCACAGCT	295
1928	commons	Sense
		Strand
KHK-1880-	Hs-Mf	25 mer	UGUGCAGACUCUAUUCCCACAGCTC	296
1929	commons	Sense
		Strand
KHK-1900-	Hs-Mf	25 mer	AGCUCAGAAGCUGGGAGUCCACACC	297
1949	commons	Sense
		Strand
KHK-1905-	Hs-Mf	25 mer	AGAAGCUGGGAGUCCACACCGCUGA	298
1954	commons	Sense
		Strand
KHK-1971-	Hs-Mf	25 mer	CUGCCCUGCCCACCAGCCUGUGATT	299
2025	commons	Sense
		Strand
KHK-1974-	Hs-Mf	25 mer	CCCUGCCCACCAGCCUGUGAUUUGA	300
2028	commons	Sense
		Strand
KHK-1975-	Hs-Mf	25 mer	CCUGCCCACCAGCCUGUGAUUUGAT	301
2029	commons	Sense
		Strand
KHK-1976-	Hs-Mf	25 mer	CUGCCCACCAGCCUGUGAUUUGATG	302
2030	commons	Sense
		Strand
KHK-1978-	Hs-Mf	25 mer	GCCCACCAGCCUGUGAUUUGAUGGG	303
2032	commons	Sense
		Strand
KHK-1979-	Hs-Mf	25 mer	CCCACCAGCCUGUGAUUUGAUGGGG	304
2033	commons	Sense
		Strand
KHK-2032-	Hs-Mf	25 mer	GCUGACUGCCCCAGAGCCUGAAAGT	305
2086	commons	Sense
		Strand
KHK-2035-	Hs-Mf	25 mer	GACUGCCCCAGAGCCUGAAAGUCTC	306
2089	commons	Sense
		Strand
KHK-2036-	Hs-Mf	25 mer	ACUGCCCCAGAGCCUGAAAGUCUCA	307
2090	commons	Sense
		Strand
KHK-2037-	Hs-Mf	25 mer	CUGCCCCAGAGCCUGAAAGUCUCAC	308
2091	commons	Sense
		Strand
KHK-2038-	Hs-Mf	25 mer	UGCCCCAGAGCCUGAAAGUCUCACC	309
2092	commons	Sense
		Strand
KHK-2039-	Hs-Mf	25 mer	GCCCCAGAGCCUGAAAGUCUCACCC	310
2093	commons	Sense
		Strand
KHK-2040-	Hs-Mf	25 mer	CCCCAGAGCCUGAAAGUCUCACCCT	311
2094	commons	Sense
		Strand
KHK-2041-	Hs-Mf	25 mer	CCCAGAGCCUGAAAGUCUCACCCTT	312
2095	commons	Sense
		Strand
KHK-2042-	Hs-Mf	25 mer	CCAGAGCCUGAAAGUCUCACCCUTG	313
2096	commons	Sense
		Strand
KHK-2043-	Hs-Mf	25 mer	CAGAGGCUGAAAGUCUCACCCUUGG	314
2097	commons	Sense
		Strand
KHK-2044-	Hs-Mf	25 mer	AGAGCCUGAAAGUCUCACCCUUGGA	315
2098	commons	Sense
		Strand
KHK-2045-	Hs-Mf	25 mer	GAGCCUGAAAGUCUCACCCUUGGAG	316
2099	commons	Sense
		Strand
KHK-2067-	Hs-Mf	25 mer	GAGCCCACCUUGGAAUUAAGGGCGT	317
2121	commons	Sense
		Strand
KHK-2069-	Hs-Mf	25 mer	GCCCACCUUGGAAUUAAGGGCGUGC	318
2123	commons	Sense
		Strand
KHK-2091-	Hs-Mf	25 mer	UGCCUCAGCCACAAAUGUGACCCAG	319
2145	commons	Sense
		Strand
KHK-2092-	Hs-Mf	25 mer	GCCUCAGCCACAAAUGUGACCCAGG	320
2146	commons	Sense
		Strand
KHK-2093-	Hs-Mf	25 mer	CCUCAGCCACAAAUGUGACCCAGGA	321
2147	commons	Sense
		Strand
KHK-2094-	Hs-Mf	25 mer	CUCAGCCACAAAUGUGACCCAGGAT	322
2148	commons	Sense
		Strand
KHK-2095-	Hs-Mf	25 mer	UCAGCCACAAAUGUGACCCAGGATA	323
2149	commons	Sense
		Strand
KHK-2096-	Hs-Mf	25 mer	CAGCCACAAAUGUGACCCAGGAUAC	324
2150	commons	Sense
		Strand
KHK-2105	Hs	25 mer	AUGUGACCCAGGAUACAGAGUGUTG	325
	unique	Sense
		Strand
KHK-2148-	Hs-Mf	25 mer	GAUCUGGAACACAUAUUGGAAUUGG	326
2197	commons	Sense
		Strand
KHK-2149-	Hs-Mf	25 mer	AUCUGGAACACAUAUUGGAAUUGGG	327
2198	commons	Sense
		Strand
KHK-2150-	Hs-Mf	25 mer	UCUGGAACACAUAUUGGAAUUGGGG	328
2199	commons	Sense
		Strand
KHK-2151-	Hs-Mf	25 mer	CUGGAACACAUAUUGGAAUUGGGGC	329
2200	commons	Sense
		Strand
KHK-2152-	Hs-Mf	25 mer	UGGAACACAUAUUGGAAUUGGGGCC	330
2201	commons	Sense
		Strand
KHK-2153-	Hs-Mf	25 mer	GGAACACAUAUUGGAAUUGGGGCCA	331
2202	commons	Sense
		Strand
KHK-2154-	Hs-Mf	25 mer	GAACACAUAUUGGAAUUGGGGCCAA	332
2203	commons	Sense
		Strand
KHK-2155-	Hs-Mf	25 mer	AACACAUAUUGGAAUUGGGGCCAAC	333
2204	commons	Sense
		Strand
KHK-2156-	Hs-Mf	25 mer	ACACAUAUUGGAAUUGGGGCCAACT	334
2205	commons	Sense
		Strand
KHK-2157-	Hs-Mf	25 mer	CACAUAUUGGAAUUGGGGCCAACTC	335
2206	commons	Sense
		Strand
KHK-2159-	Hs-Mf	25 mer	CAUAUUGGAAUUGGGGCCAACUCCA	336
2208	commons	Sense
		Strand
KHK-2160-	Hs-Mf	25 mer	AUAUUGGAAUUGGGGCCAACUCCAA	337
2209	commons	Sense
		Strand
KHK-2161-	Hs-Mf	25 mer	UAUUGGAAUUGGGGCCAACUCCAAT	338
2210	commons	Sense
		Strand
KHK-2162-	Hs-Mf	25 mer	AUUGGAAUUGGGGCCAACUCCAATA	339
2211	commons	Sense
		Strand
KHK-2163-	Hs-Mf	25 mer	UUGGAAUUGGGGCCAACUCCAAUAT	340
2212	commons	Sense
		Strand
KHK-2164-	Hs-Mf	25 mer	UGGAAUUGGGGCCAACUCCAAUATA	341
2213	commons	Sense
		Strand
KHK-2165-	Hs-Mf	25 mer	GGAAUUGGGGCCAACUCCAAUAUAG	342
2214	commons	Sense
		Strand
KHK-2166-	Hs-Mf	25 mer	GAAUUGGGGCCAACUCCAAUAUAGG	343
2215	commons	Sense
		Strand
KHK-2170-	Hs-Mf	25 mer	UGGGGCCAACUCCAAUAUAGGGUGG	344
2219	commons	Sense
		Strand
KHK-2196-	Hs-Mf	25 mer	UAAGGCCUUAUAAUGUAAAGAGCAT	345
2245	commons	Sense
		Strand
KHK-2197-	Hs-Mf	25 mer	AAGGCCUUAUAAUGUAAAGAGCATA	346
2246	commons	Sense
		Strand
KHK-2198-	Hs-Mf	25 mer	AGGCCUUAUAAUGUAAAGAGCAUAT	347
2247	commons	Sense
		Strand
KHK-2199-	Hs-Mf	25 mer	GGCCUUAUAAUGUAAAGAGCAUATA	348
2248	commons	Sense
		Strand
KHK-2200-	Hs-Mf	25 mer	GCCUUAUAAUGUAAAGAGCAUAUAA	349
2249	commons	Sense
		Strand
KHK-2201-	Hs-Mf	25 mer	CCUUAUAAUGUAAAGAGCAUAUAAT	350
2250	commons	Sense
		Strand
KHK-2205	Hs	25 mer	AUAAUGUAAAGAGCAUAUAAUGUAA	351
	unique	Sense
		Strand
KHK-2238	Hs	25 mer	AGAGUGAGACAGACCUGGAUUAAAA	352
	unique	Sense
		Strand
KHK-2260-	Hs-Mf	25 mer	AAAUCUGCCAUUUAAUUAGCUGCAT	353
2309	commons	Sense
		Strand
KHK-2261-	Hs-Mf	25 mer	AAUCUGCCAUUUAAUUAGCUGCATA	354
2310	commons	Sense
		Strand
KHK-2262-	Hs-Mf	25 mer	AUCUGCCAUUUAAUUAGCUGCAUAT	355
2311	commons	Sense
		Strand
KHK-2263-	Hs-Mf	25 mer	UCUGCCAUUUAAUUAGCUGCAUATC	356
2312	commons	Sense
		Strand
KHK-2264-	Hs-Mf	25 mer	CUGCCAUUUAAUUAGCUGCAUAUCA	357
2313	commons	Sense
		Strand
KHK-2265-	Hs-Mf	25 mer	UGCCAUUUAAUUAGCUGCAUAUCAC	358
2314	commons	Sense
		Strand
KHK-2266-	Hs-Mf	25 mer	GCCAUUUAAUUAGCUGCAUAUCACC	359
2315	commons	Sense
		Strand
KHK-2299	Hs	25 mer	CAGCACUUAACGCAAUCUGCCUCAA	360
	unique	Sense
		Strand
KHK-2317-	Hs-Mf	25 mer	GCCUCAAUUUCUUCAUCUGUCAAAT	361
2366	commons	Sense
		Strand
KHK-2318-	Hs-Mf	25 mer	CCUCAAUUUCUUCAUCUGUCAAATG	362
2367	commons	Sense
		Strand
KHK-2319-	Hs-Mf	25 mer	CUCAAUUUCUUCAUCUGUCAAAUGG	363
2368	commons	Sense
		Strand
KHK-2320-	Hs-Mf	25 mer	UCAAUUUCUUCAUCUGUCAAAUGGA	364
2369	commons	Sense
		Strand
KHK-2321-	Hs-Mf	25 mer	CAAUUUCUUCAUCUGUCAAAUGGAA	365
2370	commons	Sense
		Strand
KHK-2322-	Hs-Mf	25 mer	AAUUUCUUCAUCUGUCAAAUGGAAC	366
2371	commons	Sense
		Strand
KHK-2323-	Hs-Mf	25 mer	AUUUCUUCAUCUGUCAAAUGGAACC	367
2372	commons	Sense
		Strand
KHK-2324-	Hs-Mf	25 mer	UUUCUUCAUCUGUCAAAUGGAACCA	368
2373	commons	Sense
		Strand
KHK-2325-	Hs-Mf	25 mer	UUCUUCAUCUGUCAAAUGGAACCAA	369
2374	commons	Sense
		Strand
KHK-2326-	Hs-Mf	25 mer	UGUUCAUCUGUCAAAUGGAACCAAT	370
2375	commons	Sense
		Strand
KHK-2332	Hs	25 mer	UCUGUCAAAUGGAACCAAUUCUGCT	371
	unique	Sense
		Strand
KHK-2333	Hs	25 mer	CUGUCAAAUGGAACCAAUUCUGCTT	372
	unique	Sense
		Strand
KHK-2335	Hs	25 mer	GUCAAAUGGAACCAAUUCUGCUUGG	373
	unique	Sense
		Strand
KHK-2340	Hs	25 mer	AUGGAACCAAUUCUGCUUGGCUACA	374
	unique	Sense
		Strand
KHK-2341	Hs	25 mer	UGGAACCAAUUCUGCUUGGCUACAG	375
	unique	Sense
		Strand
KHK-2346	Hs	25 mer	CCAAUUCUGCUUGGCUACAGAAUTA	376
	unique	Sense
		Strand
KHK-2352	Hs	25 mer	CUGCUUGGCUACAGAAUUAUUGUGA	377
	unique	Sense
		Strand
KHK-2358	Hs	25 mer	GGCUACAGAAUUAUUGUGAGGAUAA	378
	unique	Sense
		Strand
KHK-2359	Hs	25 mer	GCUACAGAAUUAUUGUGAGGAUAAA	379
	unique	Sense
		Strand
KHK-2360	Hs	25 mer	CUACAGAAUUAUUGUGAGGAUAAAA	380
	unique	Sense
		Strand
KHK-2361	Hs	25 mer	UACAGAAUUAUUGUGAGGAUAAAAT	381
	unique	Sense
		Strand
KHK-2362	Hs	25 mer	ACAGAAUUAUUGUGAGGAUAAAATC	382
	unique	Sense
		Strand
KHK-2363	Hs	25 mer	CAGAAUUAUUGUGAGGAUAAAAUCA	383
	unique	Sense
		Strand
KHK-2364	Hs	25 mer	AGAAUUAUUGUGAGGAUAAAAUCAT	384
	unique	Sense
		Strand
KHK-2365	Hs	25 mer	GAAUUAUUGUGAGGAUAAAAUCATA	385
	unique	Sense
		Strand
KHK-2366	Hs	25 mer	AAUUAUUGUGAGGAUAAAAUCAUAT	386
	unique	Sense
		Strand
KHK-2367	Hs	25 mer	AUUAUUGUGAGGAUAAAAUCAUATA	387
	unique	Sense
		Strand
KHK-115-	Hs-Mf	27mer	CGAGAAUGCAAAGAGAAAAUGCGCUAC	388
154	commons	Anti-sense
		Strand
KHK-116-	Hs-Mf	27mer	UCGAGAAUGCAAAGAGAAAAUGCGCUA	389
155	commons	Anti-sense
		Strand
KHK-117-	Hs-Mf	27mer	CUCGAGAAUGCAAAGAGAAAAUGCGCU	390
156	commons	Anti-sense
		Strand
KHK-118-	Hs-Mf	27mer	UCUCGAGAAUGCAAAGAGAAAAUGCGC	391
157	commons	Anti-sense
		Strand
KHK-119-	Hs-Mf	27mer	AUCUCGAGAAUGCAAAGAGAAAAUGCG	392
158	commons	Anti-sense
		Strand
KHK-120-	Hs-Mf	27mer	GAUCUGGAGAAUGCAAAGAGAAAAUGC	393
159	commons	Anti-sense
		Strand
KHK-121-	Hs-Mf	27mer	CGAUCUGGAGAAUGCAAAGAGAAAAUG	394
160	commons	Anti-sense
		Strand
KHK-122-	Hs-Mf	27mer	GCGAUCUCGAGAAUGCAAAGAGAAAAU	395
161	commons	Anti-sense
		Strand
KHK-123-	Hs-Mf	27mer	AGCGAUCUCGAGAAUGCAAAGAGAAAA	396
162	commons	Anti-sense
		Strand
KHK-124-	Hs-Mf	27mer	AAGCGAUCUCGAGAAUGCAAAGAGAAA	397
163	commons	Anti-sense
		Strand
KHK-125-	Hs-Mf	27mer	UAAGCGAUCUCGAGAAUGCAAAGAGAA	398
164	commons	Anti-sense
		Strand
KHK-126-	Hs-Mf	27mer	CUAAGCGAUCUCGAGAAUGCAAAGAGA	399
165	commons	Anti-sense
		Strand
KHK-127-	Hs-Mf	27mer	GCUAAGCGAUCUCGAGAAUGCAAAGAG	400
166	commons	Anti-sense
		Strand
KHK-128-	Hs-Mf	27mer	GGCUAAGCGAUCUCGAGAAUGCAAAGA	401
167	commons	Anti-sense
		Strand
KHK-179	Hs	27mer	UUCCUCGCUUGUCAGAUGGACUCACAG	402
	unique	Anti-sense
		Strand
KHK-181-	Hs-Mf	27mer	GUUUCCUCGCUUGUCAGAUGGACUCAC	403
220	commons	Anti-sense
		Strand
KHK-182-	Hs-Mf	27mer	AGUUUCCUCGCUUGUCAGAUGGACUCA	404
221	commons	Anti-sense
		Strand
KHK-183-	Hs-Mf	27mer	UAGUUUCCUCGCUUGUCAGAUGGACUC	405
222	commons	Anti-sense
		Strand
KHK-184-	Hs-Mf	27mer	UUAGUUUCCUCGCUUGUCAGAUGGACU	406
223	commons	Anti-sense
		Strand
KHK-185-	Hs-Mf	27mer	CUUAGUUUCCUCGCUUGUCAGAUGGAC	407
224	commons	Anti-sense
		Strand
KHK-186-	Hs-Mf	27mer	CCUUAGUUUCCUCGCUUGUCAGAUGGA	408
225	commons	Anti-sense
		Strand
KHK-187-	Hs-Mf	27mer	GCCUUAGUUUCCUCGCUUGUCAGAUGG	409
226	commons	Anti-sense
		Strand
KHK-188-	Hs-Mf	27mer	AGCCUUAGUUUCCUCGCUUGUCAGAUG	410
227	commons	Anti-sense
		Strand
KHK-431-	Hs-Mf	27mer	CCUCUUAUCCAGGAGGAUGGUGUCCCC	411
470	commons	Anti-sense
		Strand
KHK-432-	Hs-Mf	27mer	GCCUCUUAUCCAGGAGGAUGGUGUCCC	412
471	commons	Anti-sense
		Strand
KHK-433-	Hs-Mf	27mer	UGCCUCUUAUCCAGGAGGAUGGUGUCC	413
472	commons	Anti-sense
		Strand
KHK-507-	Hs-Mf-	27mer	GGAUCUGCUUCUCUUCCAUGAGGCUAC	414
545-376-	Mm-Rn	Anti-sense
218	commons	Strand
KHK-508-	Hs-Mf-	27mer	AGGAUCUGCUUCUCUUCCAUGAGGCUA	415
546-377-	Mm-Rn	Anti-sense
219	commons	Strand
KHK-509-	Hs-Mf-	27mer	CAGGAUCUGCUUCUCUUCCAUGAGGCU	416
547-378-	Mm-Rn	Anti-sense
220	commons	Strand
KHK-510-	Hs-Mf-	27mer	ACAGGAUCUGCUUCUCUUCCAUGAGGC	417
548-379-	Mm-Rn	Anti-sense
221	commons	Strand
KHK-511-	Hs-Mf-	27mer	CACAGGAUCUGCUUCUCUUCCAUGAGG	418
549-380-	Mm-Rn	Anti-sense
222	commons	Strand
KHK-512-	Hs-Mf-	27mer	GCACAGGAUCUGCUUCUCUUCCAUGAG	419
550-381-	Mm-Rn	Anti-sense
223	commons	Strand
KHK-513-	Hs-Mf-	27mer	CGCACAGGAUCUGCUUCUCUUCCAUGA	420
551-382-	Mm-Rn	Anti-sense
224	commons	Strand
KHK-514-	Hs-Mf-	27mer	AGGCACAGGAUCUGCUUCUCUUCCAUG	421
552-383-	Mm-Rn	Anti-sense
225	commons	Strand
KHK-515-	Hs-Mf-	27mer	CAGGCACAGGAUCUGCUUCUCUUCCAU	422
553-384-	Mm-Rn	Anti-sense
226	commons	Strand
KHK-516-	Hs-Mf-	27mer	CCAGGCACAGGAUCUGCUUCUCUUCCA	423
554-385-	Mm-Rn	Anti-sense
227	commons	Strand
KHK-517-	Hs-Mf-	27mer	CCCACGCACAGGAUCUGCUUCUCUUCC	424
555-386-	Mm-Rn	Anti-sense
228	commons	Strand
KHK-518-	Hs-Mf-	27mer	CCCCACGCACAGGAUCUGCUUCUCUUC	425
556-387-	Mm-Rn	Anti-sense
229	commons	Strand
KHK-520-	Hs-Mf-	27mer	AGCCCCACGCACAGGAUCUGCUUCUCU	426
558-389-	Mm-Rn	Anti-sense
231	commons	Strand
KHK-521-	Hs-Mf-	27mer	UAGCCCCACGCACAGGAUCUGCUUCUC	427
559-390-	Mm-Rn	Anti-sense
232	commons	Strand
KHK-522-	Hs-Mf-	27mer	CUAGCCCCACGCACAGGAUCUGCUUCU	428
560-391-	Mm-Rn	Anti-sense
233	commons	Strand
KHK-541-	Hs-Mf	27mer	CUGAUGACGUCCAGCACCACUAGCCCC	429
579	commons	Anti-sense
		Strand
KHK-544-	Hs-Mf	27mer	AGGCUGAUGACGUCCAGCACCACUAGC	430
582	commons	Anti-sense
		Strand
KHK-546-	Hs-Mf	27mer	CCAGGCUGAUGACGUCCAGCACCACUA	431
584	commons	Anti-sense
		Strand
KHK-547-	Hs-Mf	27mer	ACCAGGCUGAUGACGUCCAGCACCACU	432
585	commons	Anti-sense
		Strand
KHK-548-	Hs-Mf	27mer	CACCAGGCUGAUGACGUCCAGCACCAC	433
586	commons	Anti-sense
		Strand
KHK-549-	Hs-Mf	27mer	CCACCAGGCUGAUGACGUCCAGCACCA	434
587	commons	Anti-sense
		Strand
KHK-550-	Hs-Mf	27mer	UCCACCAGGCUGAUGACGUCCAGCACC	435
588	commons	Anti-sense
		Strand
KHK-551-	Hs-Mf	27mer	GUCCACCAGGCUGAUGACGUCCAGCAC	436
589	commons	Anti-sense
		Strand
KHK-552-	Hs-Mf	27mer	UGUCCACCAGGCUGAUGACGUCCAGCA	437
590	commons	Anti-sense
		Strand
KHK-553-	Hs-Mf	27mer	UUGUCCACCAGGCUGAUGACGUCCAGC	438
591	commons	Anti-sense
		Strand
KHK-554-	Hs-Mf	27mer	CUUGUCCACCAGGCUGAUGACGUCCAG	439
592	commons	Anti-sense
		Strand
KHK-555-	Hs-Mf	27mer	ACUUGUCCACCAGGCUGAUGACGUCCA	440
593	commons	Anti-sense
		Strand
KHK-556-	Hs-Mf	27mer	UACUUGUCCACCAGGCUGAUGACGUCC	441
594	commons	Anti-sense
		Strand
KHK-557-	Hs-Mf	27mer	GUACUUGUCCACCAGGCUGAUGACGUC	442
595	commons	Anti-sense
		Strand
KHK-558-	Hs-Mf	27mer	GGUACUUGUCCACCAGGCUGAUGACGU	443
596	commons	Anti-sense
		Strand
KHK-559-	Hs-Mf	27mer	GGGUACUUGUCCACCAGGCUGAUGACG	444
597	commons	Anti-sense
		Strand
KHK-560-	Hs-Mf	27mer	AGGGUACUUGUCCACCAGGCUGAUGAC	445
598	commons	Anti-sense
		Strand
KHK-561-	Hs-Mf	27mer	UAGGGUACUUGUCCACCAGGCUGAUGA	446
599	commons	Anti-sense
		Strand
KHK-562-	Hs-Mf	27mer	UUAGGGUACUUGUCCACCAGGCUGAUG	447
600	commons	Anti-sense
		Strand
KHK-563-	Hs-Mf	27mer	CUUAGGGUACUUGUCCACCAGGCUGAU	448
601	commons	Anti-sense
		Strand
KHK-564-	Hs-Mf	27mer	CCUUAGGGUACUUGUCCACCAGGCUGA	449
602	commons	Anti-sense
		Strand
KHK-565-	Hs-Mf	27mer	UCCUUAGGGUACUUGUCCACCAGGCUG	450
603	commons	Anti-sense
		Strand
KHK-566-	Hs-Mf	27mer	CUCCUUAGGGUACUUGUCCACCAGGCU	451
604	commons	Anti-sense
		Strand
KHK-567-	Hs-Mf	27mer	CCUCCUUAGGGUACUUGUCCACCAGGC	452
605	commons	Anti-sense
		Strand
KHK-568-	Hs-Mf	27mer	UCCUCCUUAGGGUACUUGUCCACCAGG	453
606	commons	Anti-sense
		Strand
KHK-569-	Hs-Mf	27mer	GUCCUCCUUAGGGUACUUGUCCACCAG	454
607	commons	Anti-sense
		Strand
KHK-570-	Hs-Mf	27mer	AGUCCUCCUUAGGGUACUUGUCCACCA	455
608	commons	Anti-sense
		Strand
KHK-571-	Hs-Mf	27mer	GAGUCCUCCUUAGGGUACUUGUCCACC	456
609	commons	Anti-sense
		Strand
KHK-572-	Hs-Mf	27mer	CGAGUCCUCCUUAGGGUACUUGUCCAC	457
610	commons	Anti-sense
		Strand
KHK-573-	Hs-Mf	27mer	CCGAGUCCUCCUUAGGGUACUUGUCCA	458
611	commons	Anti-sense
		Strand
KHK-574-	Hs-Mf	27mer	UCCGAGUCCUCCUUAGGGUACUUGUCC	459
612	commons	Anti-sense
		Strand
KHK-575-	Hs-Mf	27mer	CUCCGAGUCCUCCUUAGGGUACUUGUC	460
613	commons	Anti-sense
		Strand
KHK-576-	Hs-Mf	27mer	UCUCCGAGUCCUCCUUAGGGUACUUGU	461
614	commons	Anti-sense
		Strand
KHK-577-	Hs-Mf	27mer	AUCUCCGAGUCCUCCUUAGGGUACUUG	462
615	commons	Anti-sense
		Strand
KHK-638-	Hs-Mf	27mer	GAGAACGGUGCAGGAGUUGGACGCGUU	463
676	commons	Anti-sense
		Strand
KHK-641-	Hs-Mf	27mer	GGAGAGAACGGUGCAGGAGUUGGACGC	464
679	commons	Anti-sense
		Strand
KHK-642-	Hs-Mf	27mer	GGGAGAGAACGGUGCAGGAGUUGGACG	465
680	commons	Anti-sense
		Strand
KHK-643-	Hs-Mf	27mer	AGGGAGAGAACGGUGCAGGAGUUGGAC	466
681	commons	Anti-sense
		Strand
KHK-644-	Hs-Mf	27mer	CAGGGAGAGAACGGUGCAGGAGUUGGA	467
682	commons	Anti-sense
		Strand
KHK-645-	Hs-Mf	27mer	GCAGGGAGAGAACGGUGCAGGAGUUGG	468
683	commons	Anti-sense
		Strand
KHK-646-	Hs-Mf	27mer	AGCAGGGAGAGAACGGUGCAGGAGUUG	469
684	commons	Anti-sense
		Strand
KHK-647-	Hs-Mf	27mer	GAGCAGGGAGAGAACGGUGCAGGAGUU	470
685	commons	Anti-sense
		Strand
KHK-650-	Hs-Mf	27mer	UCCGAGCAGGGAGAGAACGGUGCAGGA	471
688	commons	Anti-sense
		Strand
KHK-676-	Hs-Mf	27mer	AUUGAGCCCAUGAAGGCACAGGGGGCU	472
714	commons	Anti-sense
		Strand
KHK-713-	Hs-Mf	27mer	GUCGGCCACCAGGAAGUCAGCAACAUG	473
722	commons	Anti-sense
		Strand
KHK-826-	Hs-Mf	27mer	UGGAGCACAAUGGUACGGUUGCCAUUG	474
835	commons	Anti-sense
		Strand
KHK-827-	Hs-Mf	27mer	AUGGAGCACAAUGGUACGGUUGCCAUU	475
836	commons	Anti-sense
		Strand
KHK-829-	Hs-Mf	27mer	UCAUGGAGCACAAUGGUACGGUUGCCA	476
838	commons	Anti-sense
		Strand
KHK-830-	Hs-Mf	27mer	GUCAUGGAGCACAAUGGUACGGUUGCC	477
839	commons	Anti-sense
		Strand
KHK-831-	Hs-Mf	27mer	UGUCAUGGAGCACAAUGGUACGGUUGC	478
840	commons	Anti-sense
		Strand
KHK-832-	Hs-Mf	27mer	GUGUCAUGGAGCACAAUGGUACGGUUG	479
841	commons	Anti-sense
		Strand
KHK-857-	Hs-Mf	27mer	UGUAGCAGACACAUCUGGCAGGCUCGU	480
895	commons	Anti-sense
		Strand
KHK-858-	Hs-Mf	27mer	CUGUAGCAGACACAUCUGGCAGGCUCG	481
896	commons	Anti-sense
		Strand
KHK-859-	Hs-Mf	27mer	UCUGUAGCAGACACAUCUGGCAGGCUC	482
897	commons	Anti-sense
		Strand
KHK-860-	Hs-Mf-	27mer	GUCUGUAGCAGACACAUCUGGCAGGCU	483
898-729-	Mm-Rn	Anti-sense
571	commons	Strand
KHK-861-	Hs-Mf-	27mer	AGUCUGUAGCAGACACAUCUGGCAGGC	484
899-730-	Mm-Rn	Anti-sense
572	commons	Strand
KHK-862-	Hs-Mf	27mer	AAGUCUGUAGCAGACACAUCUGGCAGG	485
900	commons	Anti-sense
		Strand
KHK-865	Hs	27mer	UCAAAGUCUGUAGCAGACACAUCUGGC	486
	unique	Anti-sense
		Strand
KHK-880	Hs	27mer	GUCAGAUCAACCUUCUCAAAGUCUGUA	487
	unique	Anti-sense
		Strand
KHK-882-	Hs-Mf	27mer	GGGUCAGAUCAACCUUCUCAAAGUCUG	488
920	commons	Anti-sense
		Strand
KHK-883-	Hs-Mf	27mer	UGGGUCAGAUCAACCUUCUCAAAGUCU	489
921	commons	Anti-sense
		Strand
KHK-884-	Hs-Mf	27mer	CUGGGUCAGAUCAACCUUCUCAAAGUC	490
922	commons	Anti-sense
		Strand
KHK-885-	Hs-Mf	27mer	ACUGGGUCAGAUCAACCUUCUCAAAGU	491
923	commons	Anti-sense
		Strand
KHK-886-	Hs-Mf	27mer	AACUGGGUCAGAUCAACCUUCUCAAAG	492
924	commons	Anti-sense
		Strand
KHK-887-	Hs-Mf	27mer	GAACUGGGUCAGAUCAACCUUCUCAAA	493
925	commons	Anti-sense
		Strand
KHK-888-	Hs-Mf	27mer	UGAACUGGGUCAGAUCAACCUUCUCAA	494
926	commons	Anti-sense
		Strand
KHK-889-	Hs-Mf	27mer	UUGAACUGGGUCAGAUCAACCUUCUCA	495
927	commons	Anti-sense
		Strand
KHK-890-	Hs-Mf	27mer	CUUGAACUGGGUCAGAUCAACCUUCUC	496
928	commons	Anti-sense
		Strand
KHK-891-	Hs-Mf	27mer	ACUUGAACUGGGUCAGAUCAACCUUCU	497
929	commons	Anti-sense
		Strand
KHK-892-	Hs-Mf	27mer	CACUUGAACUGGGUCAGAUCAACCUUC	498
930	commons	Anti-sense
		Strand
KHK-893-	Hs-Mf	27mer	CCACUUGAACUGGGUCAGAUCAACCUU	499
931	commons	Anti-sense
		Strand
KHK-894-	Hs-Mf	27mer	UCCACUUGAACUGGGUCAGAUCAACCU	500
932	commons	Anti-sense
		Strand
KHK-895-	Hs-Mf	27mer	AUCCACUUGAACUGGGUCAGAUCAACC	501
933	commons	Anti-sense
		Strand
KHK-896-	Hs-Mf	27mer	GAUCCACUUGAACUGGGUCAGAUCAAC	502
934	commons	Anti-sense
		Strand
KHK-897-	Hs-Mf	27mer	GGAUCCACUUGAACUGGGUCAGAUCAA	503
935	commons	Anti-sense
		Strand
KHK-898-	Hs-Mf	27mer	UGGAUCCACUUGAACUGGGUCAGAUCA	504
936	commons	Anti-sense
		Strand
KHK-899-	Hs-Mf	27mer	GUGGAUCCACUUGAACUGGGUCAGAUC	505
937	commons	Anti-sense
		Strand
KHK-900-	Hs-Mf	27mer	UGUGGAUCCACUUGAACUGGGUCAGAU	506
938	commons	Anti-sense
		Strand
KHK-901-	Hs-Mf	27mer	AUGUGGAUCCACUUGAACUGGGUCAGA	507
939	commons	Anti-sense
		Strand
KHK-902-	Hs-Mf	27mer	AAUGUGGAUCCACUUGAACUGGGUCAG	508
940	commons	Anti-sense
		Strand
KHK-903-	Hs-Mf	27mer	CAAUGUGGAUCCACUUGAACUGGGUCA	509
941	commons	Anti-sense
		Strand
KHK-904-	Hs-Mf	27mer	UCAAUGUGGAUCCACUUGAACUGGGUC	510
942	commons	Anti-sense
		Strand
KHK-905-	Hs-Mf	27mer	CUCAAUGUGGAUCCACUUGAACUGGGU	511
943	commons	Anti-sense
		Strand
KHK-906-	Hs-Mf	27mer	CCUCAAUGUGGAUCCACUUGAACUGGG	512
944	commons	Anti-sense
		Strand
KHK-907-	Hs-Mf	27mer	CCCUCAAUGUGGAUCCACUUGAACUGG	513
945	commons	Anti-sense
		Strand
KHK-908-	Hs-Mf-	27mer	GCCCUCAAUGUGGAUCCACUUGAACUG	514
946-777-	Mm-Rn	Anti-sense
619	commons	Strand
KHK-909-	Hs-Mf-	27mer	GGCCCUCAAUGUGGAUCCACUUGAACU	515
947-778-	Mm-Rn	Anti-sense
620	commons	Strand
KHK-910-	Hs-Mf-	27mer	CGGCCCUCAAUGUGGAUCCACUUGAAC	516
948-779-	Mm-Rn	Anti-sense
621	commons	Strand
KHK-911-	Hs-Mf-	27mer	CCGGCCCUCAAUGUGGAUCCACUUGAA	517
949-780-	Mm-Rn	Anti-sense
622	commons	Strand
KHK-912-	Hs-Mf-	27mer	UCCGGCCCUCAAUGUGGAUCCACUUGA	518
950-781-	Mm-Rn	Anti-sense
623	commons	Strand
KHK-913-	Hs-Mf-	27mer	UUCCGGCCCUCAAUGUGGAUCCACUUG	519
951-782-	Mm-Rn	Anti-sense
624	commons	Strand
KHK-914-	Hs-Mf-	27mer	GUUCCGGCCCUCAAUGUGGAUCCACUU	520
952-783-	Mm-Rn	Anti-sense
625	commons	Strand
KHK-939-	Hs-Mf	27mer	GCAGCAUCUUCACCUGCUCCGAUGCGU	521
977	commons	Anti-sense
		Strand
KHK-940-	Hs-Mf	27mer	UGCAGCAUCUUCACCUGCUCCGAUGCG	522
978	commons	Anti-sense
		Strand
KHK-941-	Hs-Mf	27mer	CUGCAGCAUCUUCACCUGCUCCGAUGC	523
979	commons	Anti-sense
		Strand
KHK-942-	Hs-Mf	27mer	GCUGCAGCAUCUUCACCUGCUCCGAUG	524
980	commons	Anti-sense
		Strand
KHK-943-	Hs-Mf	27mer	CGCUGCAGCAUCUUCACCUGCUCCGAU	525
981	commons	Anti-sense
		Strand
KHK-944-	Hs-Mf	27mer	CCGCUGCAGCAUCUUCACCUGCUCCGA	526
982	commons	Anti-sense
		Strand
KHK-945-	Hs-Mf	27mer	UCCGCUGCAGCAUCUUCACCUGCUCCG	527
983	commons	Anti-sense
		Strand
KHK-946-	Hs-Mf	27mer	AUCCGCUGCAGCAUCUUCACCUGCUCC	528
984	commons	Anti-sense
		Strand
KHK-947-	Hs-Mf	27mer	UAUCCGCUGCAGCAUCUUCACCUGCUC	529
985	commons	Anti-sense
		Strand
KHK-948-	Hs-Mf-	27mer	CUAUCCGCUGCAGCAUCUUCACCUGCU	530
986-817	Mm	Anti-sense
	commons	Strand
KHK-949-	Hs-Mf-	27mer	UCUAUCCGCUGCAGCAUCUUCACCUGC	531
987-818	Mm	Anti-sense
	commons	Strand
KHK-950-	Hs-Mf-	27mer	GUCUAUCCGCUGCAGCAUCUUCACCUG	532
988-819	Mm	Anti-sense
	commons	Strand
KHK-951-	Hs-Mf-	27mer	CGUCUAUCCGCUGCAGCAUCUUCACCU	533
989-820	Mm	Anti-sense
	commons	Strand
KHK-952-	Hs-Mf-	27mer	GCGUCUAUCCGCUGCAGCAUCUUCACC	534
990-821	Mm	Anti-sense
	commons	Strand
KHK-953-	Hs-Mf-	27mer	UGCGUCUAUCCGCUGCAGCAUCUUCAC	535
991-822	Mm	Anti-sense
	commons	Strand
KHK-954-	Hs-Mf-	27mer	GUGCGUCUAUCCGCUGCAGCAUCUUCA	536
992-823	Mm	Anti-sense
	commons	Strand
KHK-955-	Hs-Mf-	27mer	UGUGCGUCUAUCCGCUGCAGCAUCUUC	537
993-824	Mm	Anti-sense
	commons	Strand
KHK-956-	Hs-Mf	27mer	GUGUGCGUCUAUCCGCUGCAGCAUCUU	538
994	commons	Anti-sense
		Strand
KHK-957-	Hs-Mf	27mer	UGUGUGCGUCUAUCCGCUGCAGCAUCU	539
995	commons	Anti-sense
		Strand
KHK-958-	Hs-Mf	27mer	UUGUGUGCGUCUAUCCGCUGCAGCAUC	540
996	commons	Anti-sense
		Strand
KHK-978-	Hs-Mf	27mer	GCUCUGGAGGCUGCCUGGUGUUGUGUG	541
1016	commons	Anti-sense
		Strand
KHK-982-	Hs-Mf	27mer	UUCUGCUCUGGAGGCUGCCUGGUGUUG	542
1020	commons	Anti-sense
		Strand
KHK-983-	Hs-Mf	27mer	CUUCUGCUCUGGAGGCUGCCUGGUGUU	543
1021	commons	Anti-sense
		Strand
KHK-984-	Hs-Mf	27mer	UCUUCUGCUCUGGAGGCUGCCUGGUGU	544
1022	commons	Anti-sense
		Strand
KHK-985-	Hs-Mf	27mer	AUCUUCUGCUCUGGAGGCUGCCUGGUG	545
1023	commons	Anti-sense
		Strand
KHK-991-	Hs-Mf	27mer	ACCCGGAUCUUCUGCUCUGGAGGCUGC	546
1029	commons	Anti-sense
		Strand
KHK-992-	Hs-Mf	27mer	CACCCGGAUCUUCUGCUCUGGAGGCUG	547
1030	commons	Anti-sense
		Strand
KHK-993-	Hs-Mf	27mer	ACACCCGGAUCUUCUGCUCUGGAGGCU	548
1031	commons	Anti-sense
		Strand
KHK-999-	Hs-Mf	27mer	CCACGGACACCCGGAUCUUCUGCUCUG	549
1037	commons	Anti-sense
		Strand
KHK-1000-	Hs-Mf	27mer	UCCACGGACACCCGGAUCUUCUGCUCU	550
1038	commons	Anti-sense
		Strand
KHK-1019-	Hs-Mf	27mer	CUCUCGUGGCUUCUCCACCUCCACGGA	551
1057	commons	Anti-sense
		Strand
KHK-1054-	Hs-Mf	27mer	ACCACGUCUCCGUAGCCAAACAGCUGG	552
1092	commons	Anti-sense
		Strand
KHK-1055-	Hs-Mf	27mer	CACCACGUCUCCGUAGCCAAACAGCUG	553
1093	commons	Anti-sense
		Strand
KHK-1057-	Hs-Mf	27mer	AACACCACGUCUCCGUAGCCAAACAGC	554
1095	commons	Anti-sense
		Strand
KHK-1058-	Hs-Mf	27mer	AAACACCACGUCUCCGUAGCCAAACAG	555
1096	commons	Anti-sense
		Strand
KHK-1059-	Hs-Mf	27mer	CAAACACCACGUCUCCGUAGCCAAACA	556
1097	commons	Anti-sense
		Strand
KHK-1060-	Hs-Mf	27mer	ACAAACACCACGUCUCCGUAGCCAAAC	557
1098	commons	Anti-sense
		Strand
KHK-1061-	Hs-Mf	27mer	GACAAACACCACGUCUCCGUAGCCAAA	558
1099	commons	Anti-sense
		Strand
KHK-1062-	Hs-Mf	27mer	UGACAAACACCACGUCUCCGUAGCCAA	559
1100	commons	Anti-sense
		Strand
KHK-1063-	Hs-Mf	27mer	CUGACAAACACCACGUCUCCGUAGCCA	560
1101	commons	Anti-sense
		Strand
KHK-1064-	Hs-Mf	27mer	GCUGACAAACACCACGUCUCCGUAGCC	561
1102	commons	Anti-sense
		Strand
KHK-1065-	Hs-Mf	27mer	UGCUGACAAACACCACGUCUCCGUAGC	562
1103	commons	Anti-sense
		Strand
KHK-1066-	Hs-Mf	27mer	UUGCUGACAAACACCACGUCUCCGUAG	563
1104	commons	Anti-sense
		Strand
KHK-1067-	Hs-Mf	27mer	UUUGCUGACAAACACCACGUCUCCGUA	564
1105	commons	Anti-sense
		Strand
KHK-1068-	Hs-Mf	27mer	CUUUGCUGACAAACACCACGUCUCCGU	565
1106	commons	Anti-sense
		Strand
KHK-1069-	Hs-Mf	27mer	UCUUUGCUGACAAACACCACGUCUCCG	566
1107	commons	Anti-sense
		Strand
KHK-1070-	Hs-Mf	27mer	AUCUUUGCUGACAAACACCACGUCUCC	567
1108	commons	Anti-sense
		Strand
KHK-1071-	Hs-Mf	27mer	CAUCUUUGCUGACAAACACCACGUCUC	568
1109	commons	Anti-sense
		Strand
KHK-1072-	Hs-Mf	27mer	ACAUCUUUGCUGACAAACACCACGUCU	569
1110	commons	Anti-sense
		Strand
KHK-1073-	Hs-Mf	27mer	CACAUCUUUGCUGACAAACACCACGUC	570
1111	commons	Anti-sense
		Strand
KHK-1074-	Hs-Mf-	27mer	CCACAUCUUUGCUGACAAACACCACGU	571
1112-943-	Mm-Rn	Anti-sense
785	commons	Strand
KHK-1075-	Hs-Mf-	27mer	GCCACAUCUUUGCUGACAAACACCACG	572
1113-944-	Mm-Rn	Anti-sense
786	commons	Strand
KHK-1076-	Hs-Mf-	27mer	GGCCACAUCUUUGCUGACAAACACCAC	573
1114-945-	Mm-Rn	Anti-sense
787	commons	Strand
KHK-1077-	Hs-Mf-	27mer	UGGCCACAUCUUUGCUGACAAACACCA	574
1115-946-	Mm-Rn	Anti-sense
788	commons	Strand
KHK-1078-	Hs-Mf-	27mer	UUGGCCACAUCUUUGCUGACAAACACC	575
1116-947-	Mm-Rn	Anti-sense
789	commons	Strand
KHK-1079-	Hs-Mf-	27mer	CUUGGCCACAUCUUUGCUGACAAACAC	576
1117-948-	Mm-Rn	Anti-sense
790	commons	Strand
KHK-1080-	Hs-Mf-	27mer	GCUUGGCCACAUCUUUGCUGACAAACA	577
1118-949-	Mm-Rn	Anti-sense
791	commons	Strand
KHK-1081-	Hs-Mf-	27mer	UGCUUGGCCACAUCUUUGCUGACAAAC	578
1119-950-	Mm-Rn	Anti-sense
792	commons	Strand
KHK-1082-	Hs-Mf-	27mer	GUGCUUGGCCACAUCUUUGCUGACAAA	579
1120-951-	Mm-Rn	Anti-sense
793	commons	Strand
KHK-1083-	Hs-Mf-	27mer	AGUGCUUGGCCACAUCUUUGCUGACAA	580
1121-952-	Mm-Rn	Anti-sense
794	commons	Strand
KHK-1084-	Hs-Mf-	27mer	AAGUGCUUGGCCACAUCUUUGCUGACA	581
1122-953-	Mm-Rn	Anti-sense
795	commons	Strand
KHK-1085-	Hs-Mf-	27mer	CAAGUGCUUGGCCACAUCUUUGCUGAC	582
1123-954-	Mm-Rn	Anti-sense
796	commons	Strand
KHK-1086-	Hs-Mf-	27mer	CCAAGUGCUUGGCCACAUCUUUGCUGA	583
1124-955-	Mm-Rn	Anti-sense
797	commons	Strand
KHK-1087-	Hs-Mf-	27mer	CCCAAGUGCUUGGCCACAUCUUUGCUG	584
1125-956-	Mm-Rn	Anti-sense
798	commons	Strand
KHK-1090-	Hs-Mf	27mer	AACCCCAAGUGCUUGGCCACAUCUUUG	585
1128	commons	Anti-sense
		Strand
KHK-1091-	Hs-Mf	27mer	GAACCCCAAGUGCUUGGCCACAUCUUU	586
1129	commons	Anti-sense
		Strand
KHK-1092-	Hs-Mf	27mer	GGAACCCCAAGUGCUUGGCCACAUCUU	587
1130	commons	Anti-sense
		Strand
KHK-1093-	Hs-Mf	27mer	UGGAACCCCAAGUGCUUGGCCACAUCU	588
1131	commons	Anti-sense
		Strand
KHK-1095-	Hs-Mf	27mer	ACUGGAACCCCAAGUGCUUGGCCACAU	589
1133	commons	Anti-sense
		Strand
KHK-1096-	Hs-Mf	27mer	GACUGGAACCCCAAGUGCUUGGCCACA	590
1134	commons	Anti-sense
		Strand
KHK-1097-	Hs-Mf	27mer	UGACUGGAACCCCAAGUGCUUGGCCAC	591
1135	commons	Anti-sense
		Strand
KHK-1099-	Hs-Mf	27mer	GCUGACUGGAACCCCAAGUGCUUGGCC	592
1137	commons	Anti-sense
		Strand
KHK-1100-	Hs-Mf	27mer	UGCUGACUGGAACCCCAAGUGCUUGGC	593
1138	commons	Anti-sense
		Strand
KHK-1101-	Hs-Mf	27mer	CUGCUGACUGGAACCCCAAGUGCUUGG	594
1139	commons	Anti-sense
		Strand
KHK-1102-	Hs-Mf	27mer	UCUGCUGACUGGAACCCCAAGUGCUUG	595
1140	commons	Anti-sense
		Strand
KHK-1103-	Hs-Mf	27mer	CUCUGCUGACUGGAACCCCAAGUGCUU	596
1141	commons	Anti-sense
		Strand
KHK-1104-	Hs-Mf	27mer	CCUCUGCUGACUGGAACCCCAAGUGCU	597
1142	commons	Anti-sense
		Strand
KHK-1106-	Hs-Mf	27mer	UUCCUCUGCUGACUGGAACCCCAAGUG	598
1144	commons	Anti-sense
		Strand
KHK-1107-	Hs-Mf	27mer	CUUCCUCUGCUGACUGGAACCCCAAGU	599
1145	commons	Anti-sense
		Strand
KHK-1135-	Hs-Mf	27mer	CUCACACGACCAUACAAGCCCCUCAAG	600
1173	commons	Anti-sense
		Strand
KHK-1136-	Hs-Mf	27mer	CCUCACACGACCAUACAAGCCCCUCAA	601
ll74	commons	Anti-sense
		Strand
KHK-1137-	Hs-Mf	27mer	UCCUCACACGACCAUACAAGCCCCUCA	602
1175	commons	Anti-sense
		Strand
KHK-1138-	Hs-Mf	27mer	UUCCUCACACGACCAUACAAGCCCCUC	603
1176	commons	Anti-sense
		Strand
KHK-1139-	Hs-Mf	27mer	UUUCCUCACACGACCAUACAAGCCCCU	604
1177	commons	Anti-sense
		Strand
KHK-1140-	Hs-Mf	27mer	CUUUCCUCACACGACCAUACAAGCCCC	605
1178	commons	Anti-sense
		Strand
KHK-1141-	Hs-Mf	27mer	CCUUUCCUCACACGACCAUACAAGCCC	606
1179	commons	Anti-sense
		Strand
KHK-1142-	Hs-Mf	27mer	CCCUUUCCUCACACGACCAUACAAGCC	607
1180	commons	Anti-sense
		Strand
KHK-1143-	Hs-Mf	27mer	CCCCUUUCCUCACACGACCAUACAAGC	608
1181	commons	Anti-sense
		Strand
KHK-1144-	Hs-Mf	27mer	GCCCCUUUCCUCACACGACCAUACAAG	609
1182	commons	Anti-sense
		Strand
KHK-1145-	Hs-Mf	27mer	AGCCCCUUUCCUCACACGACCAUACAA	610
1183	commons	Anti-sense
		Strand
KHK-1146-	Hs-Mf	27mer	CAGCCCCUUUCCUCACACGACCAUACA	611
1184	commons	Anti-sense
		Strand
KHK-1147-	Hs-Mf	27mer	ACAGCCCCUUUCCUCACACGACCAUAC	612
1185	commons	Anti-sense
		Strand
KHK-1148-	Hs-Mf	27mer	CACAGCCCCUUUCCUCACACGACCAUA	613
1186	commons	Anti-sense
		Strand
KHK-1149-	Hs-Mf	27mer	GCACAGCCCCUUUCCUCACACGACCAU	614
1187	commons	Anti-sense
		Strand
KHK-1153-	Hs-Mf	27mer	ACAAGCACAGCCCCUUUCCUCACACGA	615
1191	commons	Anti-sense
		Strand
KHK-1154-	Hs-Mf	27mer	GACAAGCACAGCCCCUUUCCUCACACG	616
1192	commons	Anti-sense
		Strand
KHK-1157-	Hs-Mf	27mer	ACAGACAAGCACAGCCCCUUUCCUCAC	617
1195	commons	Anti-sense
		Strand
KHK-1158-	Hs-Mf	27mer	CACAGACAAGCACAGCCCCUUUCCUCA	618
1196	commons	Anti-sense
		Strand
KHK-1159-	Hs-Mf	27mer	GCACAGACAAGCACAGCCCCUUUCCUC	619
1197	commons	Anti-sense
		Strand
KHK-1161-	Hs-Mf	27mer	AGGCACAGACAAGCACAGCCCCUUUCC	620
1199	commons	Anti-sense
		Strand
KHK-1163-	Hs-Mf	27mer	CCAGGCACAGACAAGCACAGCCCCUUU	621
1201	commons	Anti-sense
		Strand
KHK-1164-	Hs-Mf	27mer	CCCAGGCACAGACAAGCACAGCCCCUU	622
1202	commons	Anti-sense
		Strand
KHK-1232-	Hs-Mf	27mer	GGGUGGCGGGAAAGCAUCCGAGUGGAG	623
1270	commons	Anti-sense
		Strand
KHK-1278-	Hs-Mf-	27mer	CGGAGGCAUUGAAGGUGUCUCCAGCUC	624
1316-1147-	Mm-Rn	Anti-sense
989	commons	Strand
KHK-1279-	Hs-Mf-	27mer	ACGGAGGCAUUGAAGGUGUCUCCAGCU	625
1317-1148-	Mm-Rn	Anti-sense
990	commons	Strand
KHK-1280-	Hs-Mf-	27mer	GACGGAGGCAUUGAAGGUGUCUCCAGC	626
1318-1149-	Mm-Rn	Anti-sense
991	commons	Strand
KHK-1281-	Hs-Mf-	27mer	UGACGGAGGCAUUGAAGGUGUCUCCAG	627
1319-1150-	Mm-Rn	Anti-sense
992	commons	Strand
KHK-1282-	Hs-Mf-	27mer	AUGACGGAGGCAUUGAAGGUGUCUCCA	628
1320-1151-	Mm-Rn	Anti-sense
993	commons	Strand
KHK-1283-	Hs-Mf	27mer	GAUGACGGAGGCAUUGAAGGUGUCUCC	629
1321	commons	Anti-sense
		Strand
KHK-1284-	Hs-Mf	27mer	AGAUGACGGAGGCAUUGAAGGUGUCUC	630
1322	commons	Anti-sense
		Strand
KHK-1285-	Hs-Mf	27mer	AAGAUGACGGAGGCAUUGAAGGUGUCU	631
1323	commons	Anti-sense
		Strand
KHK-1286-	Hs-Mf	27mer	GAAGAUGACGGAGGCAUUGAAGGUGUC	632
1324	commons	Anti-sense
		Strand
KHK-1287-	Hs-Mf	27mer	UGAAGAUGACGGAGGCAUUGAAGGUGU	633
1325	commons	Anti-sense
		Strand
KHK-1288-	Hs-Mf	27mer	CUGAAGAUGACGGAGGCAUUGAAGGUG	634
1326	commons	Anti-sense
		Strand
KHK-1289-	Hs-Mf	27mer	GCUGAAGAUGACGGAGGCAUUGAAGGU	635
1327	commons	Anti-sense
		Strand
KHK-1290-	Hs-Mf	27mer	GGCUGAAGAUGACGGAGGCAUUGAAGG	636
1328	commons	Anti-sense
		Strand
KHK-1291-	Hs-Mf	27mer	AGGCUGAAGAUGACGGAGGCAUUGAAG	637
1329	commons	Anti-sense
		Strand
KHK-1292-	Hs-Mf	27mer	GAGGCUGAAGAUGACGGAGGCAUUGAA	638
1330	commons	Anti-sense
		Strand
KHK-1293-	Hs-Mf	27mer	AGAGGCUGAAGAUGACGGAGGCAUUGA	639
1331	commons	Anti-sense
		Strand
KHK-1294-	Hs-Mf	27mer	GAGAGGCUGAAGAUGACGGAGGCAUUG	640
1332	commons	Anti-sense
		Strand
KHK-1295-	Hs-Mf	27mer	GGAGAGGCUGAAGAUGACGGAGGCAUU	641
1333	commons	Anti-sense
		Strand
KHK-1297-	Hs-Mf	27mer	UGGGAGAGGCUGAAGAUGACGGAGGCA	642
1335	commons	Anti-sense
		Strand
KHK-1323-	Hs-Mf	27mer	UCAGUGCUUCCUGCACGCUCCUCCCCU	643
1361	commons	Anti-sense
		Strand
KHK-1325-	Hs-Mf	27mer	UCUCAGUGCUUCCUGCACGCUCCUCCC	644
1363	commons	Anti-sense
		Strand
KHK-1326-	Hs-Mf	27mer	AUCUCAGUGCUUCCUGCACGCUCCUCC	645
1364	commons	Anti-sense
		Strand
KHK-1327-	Hs-Mf	27mer	AAUCUCAGUGCUUCCUGCACGCUCCUC	646
1365	commons	Anti-sense
		Strand
KHK-1328-	Hs-Mf	27mer	GAAUCUCAGUGCUUCCUGCACGCUCCU	647
1366	commons	Anti-sense
		Strand
KHK-1329-	Hs-Mf	27mer	CGAAUCUCAGUGCUUCCUGCACGCUCC	648
1367	commons	Anti-sense
		Strand
KHK-1330-	Hs-Mf	27mer	CCGAAUCUCAGUGCUUCCUGCACGCUC	649
1368	commons	Anti-sense
		Strand
KHK-1331-	Hs-Mf	27mer	CCCGAAUCUCAGUGCUUCCUGCACGCU	650
1369	commons	Anti-sense
		Strand
KHK-1332-	Hs-Mf	27mer	ACCCGAAUCUCAGUGCUUCCUGCACGC	651
1370	commons	Anti-sense
		Strand
KHK-1333-	Hs-Mf	27mer	CACCCGAAUCUCAGUGCUUCCUGCACG	652
1371	commons	Anti-sense
		Strand
KHK-1334-	Hs-Mf	27mer	GCACCCGAAUCUCAGUGCUUCCUGCAC	653
1372	commons	Anti-sense
		Strand
KHK-1335-	Hs-Mf	27mer	GGCACCCGAAUCUCAGUGCUUCCUGCA	654
1373	commons	Anti-sense
		Strand
KHK-1336-	Hs-Mf	27mer	UGGCACCCGAAUCUCAGUGCUUCCUGC	655
1374	commons	Anti-sense
		Strand
KHK-1385-	Hs-Mf	27mer	UCACACGAUGCCAUCAAAGCCCUGCAG	656
1423	commons	Anti-sense
		Strand
KHK-1387-	Hs-Mf	27mer	UCUCACACGAUGCCAUCAAAGCCCUGC	657
1425	commons	Anti-sense
		Strand
KHK-1388-	Hs-Mf	27mer	CUCUCACACGAUGCCAUCAAAGCCCUG	658
1426	commons	Anti-sense
		Strand
KHK-1389-	Hs-Mf	27mer	GCUCUCACACGAUGCCAUCAAAGCCCU	659
1427	commons	Anti-sense
		Strand
KHK-1538-	Hs-Mf	27mer	CCUGUGGGGAACACAGGACACAGGCAG	660
1588	commons	Anti-sense
		Strand
KHK-1540-	Hs-Mf	27mer	UCCCUGUGGGGAACACAGGACACAGGC	661
1590	commons	Anti-sense
		Strand
KHK-1542-	Hs-Mf	27mer	UCUCCCUGUGGGGAACACAGGACACAG	662
1592	commons	Anti-sense
		Strand
KHK-1665-	Hs-Mf	27mer	UCGAAGAGUCAGAGCCUCAGGAAUGCC	663
1708	commons	Anti-sense
		Strand
KHK-1666-	Hs-Mf	27mer	AUCGAAGAGUCAGAGCCUCAGGAAUGC	664
1709	commons	Anti-sense
		Strand
KHK-1667-	Hs-Mf	27mer	GAUCGAAGAGUCAGAGCCUCAGGAAUG	665
1710	commons	Anti-sense
		Strand
KHK-1707-	Hs-Mf	27mer	GGCGGAGAGGUUAAUUUGGGGAAUGGA	666
1750	commons	Anti-sense
		Strand
KHK-1708-	Hs-Mf	27mer	GGGCGGAGAGGUUAAUUUGGGGAAUGG	667
1751	commons	Anti-sense
		Strand
KHK-1709-	Hs-Mf	27mer	UGGGCGGAGAGGUUAAUUUGGGGAAUG	668
1752	commons	Anti-sense
		Strand
KHK-1869-	Hs-Mf	27mer	AUAGAGUCUGCACAACGCAGGGCCCCG	669
1918	commons	Anti-sense
		Strand
KHK-1870-	Hs-Mf	27mer	AAUAGAGUCUGCACAACGCAGGGCCCC	670
1919	commons	Anti-sense
		Strand
KHK-1871-	Hs-Mf	27mer	GAAUAGAGUCUGCACAACGCAGGGCCC	671
1920	commons	Anti-sense
		Strand
KHK-1872-	Hs-Mf	27mer	GGAAUAGAGUCUGCACAACGCAGGGCC	672
1921	commons	Anti-sense
		Strand
KHK-1873-	Hs-Mf	27mer	GGGAAUAGAGUCUGCACAACGCAGGGC	673
1922	commons	Anti-sense
		Strand
KHK-1874-	Hs-Mf	27mer	UGGGAAUAGAGUCUGCACAACGCAGGG	674
1923	commons	Anti-sense
		Strand
KHK-1875-	Hs-Mf	27mer	GUGGGAAUAGAGUCUGCACAACGCAGG	675
1924	commons	Anti-sense
		Strand
KHK-1876-	Hs-Mf	27mer	UGUGGGAAUAGAGUCUGCACAACGCAG	676
1925	commons	Anti-sense
		Strand
KHK-1877-	Hs-Mf	27mer	CUGUGGGAAUAGAGUCUGCACAACGCA	677
1926	commons	Anti-sense
		Strand
KHK-1878-	Hs-Mf	27mer	GCUGUGGGAAUAGAGUCUGCACAACGC	678
1927	commons	Anti-sense
		Strand
KHK-1879-	Hs-Mf	27mer	AGCUGUGGGAAUAGAGUCUGCACAACG	679
1928	commons	Anti-sense
		Strand
KHK-1880-	Hs-Mf	27mer	GAGCUGUGGGAAUAGAGUCUGCACAAC	680
1929	commons	Anti-sense
		Strand
KHK-1900-	Hs-Mf	27mer	GGUGUGGACUCCCAGCUUCUGAGCUGU	681
1949	commons	Anti-sense
		Strand
KHK-1905-	Hs-Mf	27mer	UCAGCGGUGUGGACUCCCAGCUUCUGA	682
1954	commons	Anti-sense
		Strand
KHK-1971-	Hs-Mf	27mer	AAUCACAGGCUGGUGGGCAGGGCAGAG	683
2025	commons	Anti-sense
		Strand
KHK-1974-	Hs-Mf	27mer	UCAAAUCACAGGCUGGUGGGCAGGGCA	684
2028	commons	Anti-sense
		Strand
KHK-1975-	Hs-Mf	27mer	AUCAAAUCACAGGCUGGUGGGCAGGGC	685
2029	commons	Anti-sense
		Strand
KHK-1976-	Hs-Mf	27mer	CAUCAAAUCACAGGCUGGUGGGCAGGG	686
2030	commons	Anti-sense
		Strand
KHK-1978-	Hs-Mf	27mer	CCCAUCAAAUCACAGGCUGGUGGGCAG	687
2032	commons	Anti-sense
		Strand
KHK-1979-	Hs-Mf	27mer	CCCCAUCAAAUCACAGGCUGGUGGGCA	688
2033	commons	Anti-sense
		Strand
KHK-2032-	Hs-Mf	27mer	ACUUUCAGGCUCUGGGGCAGUCAGCGG	689
2086	commons	Anti-sense
		Strand
KHK-2035-	Hs-Mf	27mer	GAGACUUUCAGGCUCUGGGGCAGUCAG	690
2089	commons	Anti-sense
		Strand
KHK-2036-	Hs-Mf	27mer	UGAGACUUUCAGGCUCUGGGGCAGUCA	691
2090	commons	Anti-sense
		Strand
KHK-2037-	Hs-Mf	27mer	GUGAGACUUUCAGGCUCUGGGGCAGUC	692
2091	commons	Anti-sense
		Strand
KHK-2038-	Hs-Mf	27mer	GGUGAGACUUUCAGGCUCUGGGGCAGU	693
2092	commons	Anti-sense
		Strand
KHK-2039-	Hs-Mf	27mer	GGGUGAGACUUUCAGGCUCUGGGGCAG	694
2093	commons	Anti-sense
		Strand
KHK-2040-	Hs-Mf	27mer	AGGGUGAGACUUUCAGGCUCUGGGGCA	695
2094	commons	Anti-sense
		Strand
KHK-2041-	Hs-Mf	27mer	AAGGGUGAGACUUUCAGGCUCUGGGGC	696
2095	commons	Anti-sense
		Strand
KHK-2042-	Hs-Mf	27mer	CAAGGGUGAGACUUUCAGGCUCUGGGG	697
2096	commons	Anti-sense
		Strand
KHK-2043-	Hs-Mf	27mer	CCAAGGGUGAGACUUUCAGGCUCUGGG	698
2097	commons	Anti-sense
		Strand
KHK-2044-	Hs-Mf	27mer	UCCAAGGGUGAGACUUUCAGGCUCUGG	699
2098	commons	Anti-sense
		Strand
KHK-2045-	Hs-Mf	27mer	CUCCAAGGGUGAGACUUUCAGGCUCUG	700
2099	commons	Anti-sense
		Strand
KHK-2067-	Hs-Mf	27mer	ACGCCCUUAAUUCCAAGGUGGGCUCCA	701
2121	commons	Anti-sense
		Strand
KHK-2069-	Hs-Mf	27mer	GCACGCCCUUAAUUCCAAGGUGGGCUC	702
2123	commons	Anti-sense
		Strand
KHK-2091-	Hs-Mf	27mer	CUGGGUCACAUUUGUGGCUGAGGCACG	703
2145	commons	Anti-sense
		Strand
KHK-2092-	Hs-Mf	27mer	CCUGGGUCACAUUUGUGGCUGAGGCAC	704
2146	commons	Anti-sense
		Strand
KHK-2093-	Hs-Mf	27mer	UCCUGGGUCACAUUUGUGGCUGAGGCA	705
2147	commons	Anti-sense
		Strand
KHK-2094-	Hs-Mf	27mer	AUCCUGGGUCACAUUUGUGGCUGAGGC	706
2148	commons	Anti-sense
		Strand
KHK-2095-	Hs-Mf	27mer	UAUCCUGGGUCACAUUUGUGGCUGAGG	707
2149	commons	Anti-sense
		Strand
KHK-2096-	Hs-Mf	27mer	GUAUCCUGGGUCACAUUUGUGGCUGAG	708
2150	commons	Anti-sense
		Strand
KHK-2105	Hs	27mer	CAACACUCUGUAUCCUGGGUCACAUUU	709
	unique	Anti-sense
		Strand
KHK-2148-	Hs-Mf	27mer	CCAAUUCCAAUAUGUGUUCCAGAUCGG	710
2197	commons	Anti-sense
		Strand
KHK-2149-	Hs-Mf	27mer	CCCAAUUCCAAUAUGUGUUCCAGAUCG	711
2198	commons	Anti-sense
		Strand
KHK-2150-	Hs-Mf	27mer	CCCCAAUUCCAAUAUGUGUUCCAGAUC	712
2199	commons	Anti-sense
		Strand
KHK-2151-	Hs-Mf	27mer	GCCCCAAUUCCAAUAUGUGUUCCAGAU	713
2200	commons	Anti-sense
		Strand
KHK-2152-	Hs-Mf	27mer	GGCCCCAAUUCCAAUAUGUGUUCCAGA	714
2201	commons	Anti-sense
		Strand
KHK-2153-	Hs-Mf	27mer	UGGCCCCAAUUCCAAUAUGUGUUCCAG	715
2202	commons	Anti-sense
		Strand
KHK-2154-	Hs-Mf	27mer	UUGGCCCCAAUUCCAAUAUGUGUUCCA	716
2203	commons	Anti-sense
		Strand
KHK-2155-	Hs-Mf	27mer	GUUGGCCCCAAUUCCAAUAUGUGUUCC	717
2204	commons	Anti-sense
		Strand
KHK-2156-	Hs-Mf	27mer	AGUUGGCCCCAAUUCCAAUAUGUGUUC	718
2205	commons	Anti-sense
		Strand
KHK-2157-	Hs-Mf	27mer	GAGUUGGCCCCAAUUCCAAUAUGUGUU	719
2206	commons	Anti-sense
		Strand
KHK-2159-	Hs-Mf	27mer	UGGAGUUGGCCCCAAUUCCAAUAUGUG	720
2208	commons	Anti-sense
		Strand
KHK-2160-	Hs-Mf	27mer	UUGGAGUUGGCCCCAAUUCCAAUAUGU	721
2209	commons	Anti-sense
		Strand
KHK-2161-	Hs-Mf	27mer	AUUGGAGUUGGCCCCAAUUCCAAUAUG	722
2210	commons	Anti-sense
		Strand
KHK-2162-	Hs-Mf	27mer	UAUUGGAGUUGGCCCCAAUUCCAAUAU	723
2211	commons	Anti-sense
		Strand
KHK-2163-	Hs-Mf	27mer	AUAUUGGAGUUGGCCCCAAUUCCAAUA	724
2212	commons	Anti-sense
		Strand
KHK-2164-	Hs-Mf	27mer	UAUAUUGGAGUUGGCCCCAAUUCCAAU	725
2213	commons	Anti-sense
		Strand
KHK-2165-	Hs-Mf	27mer	CUAUAUUGGAGUUGGCCCCAAUUCCAA	726
2214	commons	Anti-sense
		Strand
KHK-2166-	Hs-Mf	27mer	CCUAUAUUGGAGUUGGCCCCAAUUCCA	727
2215	commons	Anti-sense
		Strand
KHK-2170-	Hs-Mf	27mer	CCACCCUAUAUUGGAGUUGGCCCCAAU	728
2219	commons	Anti-sense
		Strand
KHK-2196-	Hs-Mf	27mer	AUGCUCUUUACAUUAUAAGGCCUUACC	729
2245	commons	Anti-sense
		Strand
KHK-2197-	Hs-Mf	27mer	UAUGCUCUUUACAUUAUAAGGCCUUAC	730
2246	commons	Anti-sense
		Strand
KHK-2198-	Hs-Mf	27mer	AUAUGCUCUUUACAUUAUAAGGCCUUA	731
2247	commons	Anti-sense
		Strand
KHK-2199-	Hs-Mf	27mer	UAUAUGCUCUUUACAUUAUAAGGCCUU	732
2248	commons	Anti-sense
		Strand
KHK-2200-	Hs-Mf	27mer	UUAUAUGCUCUUUACAUUAUAAGGCCU	733
2249	commons	Anti-sense
		Strand
KHK-2201-	Hs-Mf	27mer	AUUAUAUGCUCUUUACAUUAUAAGGCC	734
2250	commons	Anti-sense
		Strand
KHK-2205	Hs	27mer	UUACAUUAUAUGCUCUUUACAUUAUAA	735
	unique	Anti-sense
		Strand
KHK-2238	Hs	27mer	UUUUAAUCCAGGUCUGUCUCACUCUAA	736
	unique	Anti-sense
		Strand
KHK-2260-	Hs-Mf	27mer	AUGCAGCUAAUUAAAUGGCAGAUUUUA	737
2309	commons	Anti-sense
		Strand
KHK-2261-	Hs-Mf	27mer	UAUGCAGCUAAUUAAAUGGCAGAUUUU	738
2310	commons	Anti-sense
		Strand
KHK-2262-	Hs-Mf	27mer	AUAUGCAGCUAAUUAAAUGGCAGAUUU	739
2311	commons	Anti-sense
		Strand
KHK-2263-	Hs-Mf	27mer	GAUAUGCAGCUAAUUAAAUGGCAGAUU	740
2312	commons	Anti-sense
		Strand
KHK-2264-	Hs-Mf	27mer	UGAUAUGCAGCUAAUUAAAUGGCAGAU	741
2313	commons	Anti-sense
		Strand
KHK-2265-	Hs-Mf	27mer	GUGAUAUGCAGCUAAUUAAAUGGCAGA	742
2314	commons	Anti-sense
		Strand
KHK-2266-	Hs-Mf	27mer	GGUGAUAUGCAGCUAAUUAAAUGGCAG	743
2315	commons	Anti-sense
		Strand
KHK-2299	Hs	27mer	UUGAGGCAGAUUGCGUUAAGUGCUGUA	744
	unique	Anti-sense
		Strand
KHK-2317-	Hs-Mf	27mer	AUUUGACAGAUGAAGAAAUUGAGGCAG	745
2366	commons	Anti-sense
		Strand
KHK-2318-	Hs-Mf	27mer	CAUUUGACAGAUGAAGAAAUUGAGGCA	746
2367	commons	Anti-sense
		Strand
KHK-2319-	Hs-Mf	27mer	CCAUUUGACAGAUGAAGAAAUUGAGGC	747
2368	commons	Anti-sense
		Strand
KHK-2320-	Hs-Mf	27mer	UCCAUUUGACAGAUGAAGAAAUUGAGG	748
2369	commons	Anti-sense
		Strand
KHK-2321-	Hs-Mf	27mer	UUCCAUUUGACAGAUGAAGAAAUUGAG	749
2370	commons	Anti-sense
		Strand
KHK-2322-	Hs-Mf	27mer	GUUCCAUUUGACAGAUGAAGAAAUUGA	750
2371	commons	Anti-sense
		Strand
KHK-2323-	Hs-Mf	27mer	GGUUCCAUUUGACAGAUGAAGAAAUUG	751
2372	commons	Anti-sense
		Strand
KHK-2324-	Hs-Mf	27mer	UGGUUCCAUUUGACAGAUGAAGAAAUU	752
2373	commons	Anti-sense
		Strand
KHK-2325-	Hs-Mf	27mer	UUGGUUCCAUUUGACAGAUGAAGAAAU	753
2374	commons	Anti-sense
		Strand
KHK-2326-	Hs-Mf	27mer	AUUGGUUCCAUUUGACAGAUGAAGAAA	754
2375	commons	Anti-sense
		Strand
KHK-2332	Hs	27mer	AGCAGAAUUGGUUCCAUUUGACAGAUG	755
	unique	Anti-sense
		Strand
KHK-2333	Hs	27mer	AAGCAGAAUUGGUUCCAUUUGACAGAU	756
	unique	Anti-sense
		Strand
KHK-2335	Hs	27mer	CCAAGCAGAAUUGGUUCCAUUUGACAG	757
	unique	Anti-sense
		Strand
KHK-2340	Hs	27mer	UGUAGCCAAGCAGAAUUGGUUCCAUUU	758
	unique	Anti-sense
		Strand
KHK-2341	Hs	27mer	CUGUAGCCAAGCAGAAUUGGUUCCAUU	759
	unique	Anti-sense
		Strand
KHK-2346	Hs	27mer	UAAUUCUGUAGCCAAGCAGAAUUGGUU	760
	unique	Anti-sense
		Strand
KHK-2352	Hs	27mer	UCACAAUAAUUCUGUAGCCAAGCAGAA	761
	unique	Anti-sense
		Strand
KHK-2358	Hs	27mer	UUAUCCUCACAAUAAUUCUGUAGCCAA	762
	unique	Anti-sense
		Strand
KHK-2359	Hs	27mer	UUUAUCCUCACAAUAAUUCUGUAGCCA	763
	unique	Anti-sense
		Strand
KHK-2360	Hs	27mer	UUUUAUCCUCACAAUAAUUCUGUAGCC	764
	unique	Anti-sense
		Strand
KHK-2361	Hs	27mer	AUUUUAUCCUCACAAUAAUUCUGUAGC	765
	unique	Anti-sense
		Strand
KHK-2362	Hs	27mer	GAUUUUAUCCUCACAAUAAUUCUGUAG	766
	unique	Anti-sense
		Strand
KHK-2363	Hs	27mer	UGAUUUUAUCCUCACAAUAAUUCUGUA	767
	unique	Anti-sense
		Strand
KHK-2364	Hs	27mer	AUGAUUUUAUCCUCACAAUAAUUCUGU	768
	unique	Anti-sense
		Strand
KHK-2365	Hs	27mer	UAUGAUUUUAUCCUCACAAUAAUUCUG	769
	unique	Anti-sense
		Strand
KHK-2366	Hs	27mer	AUAUGAUUUUAUCCUCACAAUAAUUCU	770
	unique	Anti-sense
		Strand
KHK-2367	Hs	27mer	UAUAUGAUUUUAUCCUCACAAUAAUUC	771
	unique	Anti-sense
		Strand
Forward	NHP	N/A	TGCCTTCATGGGCTCAATG	772
Primer	KHK
Reverse	NHP	N/A	TCGGCCACCAGGAAGTCA	773
Primer	KHK
KHK-510-	Hs-Mf-	Low-2′-	[mCs][mU][mC][mA][mU][mG][mG][fA]	774
548-379-	Mm-Rn	Fluoro	[fA][fG][fA][mG][mA][mA][mG][mC]
221	commons	Pattern	[mA][mG][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-516-	Hs-Mf-	Low-2′-	[mGs][mA][mA][mG][mA][mG][mA][fA]	775
554-385-	Mm-Rn	Fluoro	[fG][fC][fA][mG][mA][mU][mC][mC]
227	commons	Pattern	[mU][mG][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-829-	Hs-Mf	Low-2′-	[mGs][mC][mA][mA][mC][mC][mG][fU]	776
838	commons	Fluoro	[fA][fC][fC][mA][mU][mU][mG][mU]
		Pattern	[mG][mC][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-860-	Hs-Mf-	Low-2′-	[mCs][mC][mU][mG][mC][mC][mA][fG]	777
898-729-	Mm-Rn	Fluoro	[fA][fU][fG][mU][mG][mU][mC][mU]
571	commons	Pattern	[mG][mC][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-861-	Hs-Mf-	Low-2′-	[mCs][mU][mG][mC][mC][mA][mG][fA]	778
899-730-	Mm-Rn	Fluoro	[fU][fG][fU][mG][mU][mC][mU][mG]
572	commons	Pattern	[mC][mU][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-865	Hs	Low-2′-	[mCs][mA][mG][mA][mU][mG][mU][fG]	779
	unique	Fluoro	[fU][fC][fU][mG][mC][mU][mA][mC]
		Pattern	[mA][mG][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-882-	Hs-Mf	Low-2′-	[mGs][mA][mC][mU][mU][mU][mG][fA]	780
920	commons	Fluoro	[fG][fA][fA][mG][mG][mU][mU][mG]
		Pattern	[mA][mU][mC][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-883-	Hs-Mf	Low-2′-	[mAs][mC][mU][mU][mU][mG][mA][fG]	781
921	commons	Fluoro	[fA][fA][fG][mG][mU][mU][mG][mA]
		Pattern	[mU][mC][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-885-	Hs-Mf	Low-2′-	[mUs][mU][mU][mG][mA][mG][mA][fA]	782
923	commons	Fluoro	[fG][fG][fU][mU][mG][mA][mU][mC]
		Pattern	[mU][mG][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1054-	Hs-Mf	Low-2′-	[mAs][mG][fC][mU][mG][mU][mU][fU]	783
1092	commons	Fluoro	[fG][fG][m][fU][fA][mC][mG][mG]
		Pattern	C[fA][mG][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1075-	Hs-Mf-	Low-2′-	[mUs][mG][mG][mU][mG][mU][mU][fU]	784
1113-944-	Mm-Rn	Fluoro	[fG][fU][fC][mA][mG][mC][mA][mA]
786	commons	Pattern	[mA][mG][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1078-	Hs-Mf-	Low-2′-	[mUs][mG][mU][mU][mU][mG][mU][fC]	785
1116-947-	Mm-Rn	Fluoro	[fA][fG][fC][mA][mA][mA][mG][mA]
789	commons	Pattern	[mU][mG][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1281-	Hs-Mf-	Low-2′-	[mGs][mG][mA][mG][mA][mC][mA][fC]	786
1319-1150-	Mm-Rn	Fluoro	[fC][fU][fU][mC][mA][mA][mU][mG]
992	commons	Pattern	[mC][mC][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1288-	Hs-Mf	Low-2′-	[mCs][mC][mU][mU][mC][mA][mA][fU]	787
1326	commons	Fluoro	[fG][fC][fC][mU][mC][mC][mG][mU]
		Pattern	[mC][mA][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1290-	Hs-Mf	Low-2′-	[mUs][mU][mC][mA][mA][mU][mG][fC]	788
1328	commons	Fluoro	[fC][fU][fC][mC][mG][mU][mC][mA]
		Pattern	[mU][mC][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1148-	Hs-Mf	Low-2′-	[mUs][mG][mG][mU][mC][mG][mU][fG]	789
1186	commons	Fluoro	[fU][fG][fA][mG][mG][mA][mA][mA]
		Pattern	[mG][mG][mG][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1152-	Hs-Mf	Low-2′-	[mCs][mG][mU][mG][mU][mG][mA][fG]	790
1190	commons	Fluoro	[fG][fA][fA][mA][mG][mG][mG][mG]
		Pattern	[mC][mU][mG][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1154-	Hs-Mf	Low-2′-	[mUs][mG][mU][mG][mA][mG][mG][fA]	791
1192	commons	Fluoro	[fA][fA][fG][mG][mG][mG][mC][mU]
		Pattern	[mG][mU][mG][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1155-	Hs-Mf	Low-2′-	[mGs][mU][mG][mA][mG][mG][mA][fA]	792
1193	commons	Fluoro	[fA][fG][fG][mG][mG][mC][mU][mG]
		Pattern	[mU][mG][mC][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1277	Hs	Low-2′-	[mAs][mG][mC][mU][mG][mG][mA][fG]	793
	unique	Fluoro	[fA][fC][fA][mC][mC][mU][mU][mC]
		Pattern	[mA][mA][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1147-	Hs-Mf	Low-2′-	[mAs][mU][mG][mG][mU][mC][mG][fU]	794
1185	commons	Fluoro	[fG][fU][fG][mA][mG][mG][mA][mA]
		Pattern	[mA][mG][mG][mA][G][mC][mA][mG]
		Modified	m[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-869-	Hs-Mf	Low-2′-	[mUs][mG][mU][mG][mU][mC][mU][fG]	795
934	commons	Fluoro	[fC][fU][fA][mC][mA][mG][mA][mC]
		Pattern	[mU][mU][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-alNAc][mG][mG][mC]
		Strand	G[mU][mG][mC]
KHK-873	Hs	Low-2′-	[mUs][mC][mU][mG][mC][mU][mA][fC]	796
	unique	Fluoro	[fA][fG][fA][mC][mU][mU][mU][mG]
		Pattern	[mA][mG][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-879	Hs	Low-2′-	[mAs][mC][mA][mG][mA][mC][mU][fU]	797
	unique	Fluoro	[fU][fG][fA][mG][mA][mA][mG][mG]
		Pattern	[mU][mU][mG][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-881	Hs	Low-2′-	[mAs][mG][mA][mC][mU][mU][mU][fG]	798
	unique	Fluoro	[fA][fG][fA][mA][mG][mG][mU][mU]
		Pattern	[mG][mA][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-896-	Hs-Mf	Low-2′-	[mUs][mG][mA][mU][mC][mU][mG][fA]	799
934	commons	Fluoro	[fC][fC][fC][mA][mG][mU][mU][mC]
		Pattern	[mA][mA][mG][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1064-	Hs-Mf	Low-2′-	[mCs][mU][mA][mC][mG][mG][mA][fG]	800
1102	commons	Fluoro	[fA][fC][fG][mU][mG][mG][mU][mG]
		Pattern	[mU][mU][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1077-	Hs-Mf-	Low-2′-	[mGs][mU][mG][mU][mU][mU][mG][fU]	801
1115-946-	Mm-Rn	Fluoro	[fC][fA][fG][mC][mA][mA][mA][mG]
788	commons	Pattern	[mA][mU][mG][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1080-	Hs-Mf-	Low-2′-	[mUs][mU][mU][mG][mU][mC][mA][fG]	802
1118-949-	Mm-Rn	Fluoro	[fC][fA][fA][mA][mG][mA][mU][mG]
791	commons	Pattern	[mU][mG][mG][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-alNAc][mG][mG][mC]
		Strand	G[mU][mG][mC]
KHK-1106-	Hs-Mf	Low-2′-	[mCs][mU][mU][mG][mG][mG][mG][fU]	803
1144	commons	Fluoro	[fU][fC][fC][mA][mG][mU][mC][mA]
		Pattern	[mG][mC][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1334-	Hs-Mf	Low-2′-	[mGs][mC][mA][mG][mG][mA][mA][fG]	804
1372	commons	Fluoro	[fC][fA][fC][mU][mG][mA][mG][mA]
		Pattern	[mU][mU][mC][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-516-	Hs-Mf-	Med-2′-	[mGs][mA][fA][mG][mA][mG][mA][fA]	805
554-385-	Mm-Rn	Fluoro	[fG][fC][mA][fG][fA][mU][mC][mC]
227	commons	Pattern	[fU][mG][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-804	Hs	Med-2′-	[mUs][mG][fC][mU][mG][mC][mA][fU]	806
	unique	Fluoro	[fC][fA][mU][fC][fA][mA][mC][mA]
		Pattern	[fA][mC][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-829-	Hs-Mf	Med-2′-	[mGs][mC][fA][mA][mC][mC][mG][fU]	807
838	commons	Fluoro	[fA][fC][mC][fA][fU][mU][mG][mU]
		Pattern	[fG][mC][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-861-	Hs-Mf-	Med-2′-	[mCs][mU][fG][mC][mC][mA][mG][fA]	808
899-730-	Mm-Rn	Fluoro	[fU][fG][mU][fG][fU][mC][mU][mG]
572	commons	Pattern	[fC][mU][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-865	Hs	Med-2′-	[mCs][mA][fG][mA][mU][mG][mU][fG]	809
	unique	Fluoro	[fU][fC][mU][fG][fC][mU][mA][mC]
		Pattern	[fA][mG][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-882-	Hs-Mf	Med-2′-	[mGs][mA][fC][mU][mU][mU][mG][fA]	810
920	commons	Fluoro	[fG][fA][mA][fG][fG][mU][mU][mG]
		Pattern	[fA][mU][mC][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-883-	Hs-Mf	Med-2′-	[mAs][mC][fU][mU][mU][mG][mA][fG]	811
921	commons	Fluoro	[fA][fA][mG][fG][fU][mU][mG][mA]
		Pattern	[fU][mC][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-885-	Hs-Mf	Med-2′-	[mUs][mU][fU][mG][mA][mG][mA][fA]	812
923	commons	Fluoro	[fG][fG][mU][fU][fG][mA][mU][mC]
		Pattern	[fU][mG][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1054-	Hs-Mf	Med-2′-	[mAs][mG][fC][mU][mG][mU][mU][fU]	813
1092	commons	Fluoro	[fG][fG][mC][fU][fA][mC][mG][mG]
		Pattern	[fA][mG][mA][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1076-	Hs-Mf-	Med-2′-	[mGs][mG][fU][mG][mU][mU][mU][fG]	814
1114-945-	Mm-Rn	Fluoro	[fU][fC][mA][fG][fC][mA][mA][mA]
787	commons	Pattern	[fG][mA][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1078-	Hs-Mf-	Med-2′-	[mUs][mG][fU][mU][mU][mG][mU][fC]	815
1116-947-	Mm-Rn	Fluoro	[fA][fG][mC][fA][fA][mA][mG][mA]
789	commons	Pattern	[fU][mG][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1288-	Hs-Mf	Med-2′-	[mCs][mC][fU][mU][mC][mA][mA][fU]	816
1326	commons	Fluoro	[fG][fC][mC][fU][fC][mC][mG][mU]
		Pattern	[fC][mA][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1290-	Hs-Mf	Med-2′-	[mUs][mU][fC][mA][mA][mU][mG][fC]	817
1328	commons	Fluoro	[fC][fU][mC][fC][fG][mU][mC][mA]
		Pattern	[fU][mC][mU][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-1334-	Hs-Mf	Med-2′-	[mGs][mC][fA][mG][mG][mA][mA][fG]	818
1372	commons	Fluoro	[fC][fA][mC][fU][fG][mA][mG][mA]
		Pattern	[fU][mU][mC][mA][mG][mC][mA][mG]
		Modified	[mC][mC][mG][ademA-GalNAc][ademA-
		Sense	GalNAc][ademA-GalNAc][mG][mG][mC]
		Strand	[mU][mG][mC]
KHK-510-	Hs-Mf-	Low-2′-	[MePhosphonate-4O-	819
548-379-	Mm-Rn	Fluoro	mUs][fUs][fCs][fU][fG][mC][fU][mU]
221	commons	Pattern	[mC][fU][mC][mU][mU][fC][mC][mA]
		Modified	[mU][mG][mA][mGs][mGs][mG]
		Anti-sense
		Strand
KHK-516-	Hs-Mf-	Low-2′-	[MePhosphonate-4O-	820
554-385-	Mm-Rn	Fluoro	mUs][fAs][fC][fA][fG][mG][fA][mU]
227	commons	Pattern	[mC][fU][mG][mC][mU][fU][mC][mU]
		Modified	[mC][mU][mU][mCs][mGs][mG]
		Anti-sense
		Strand
KHK-829-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	821
838	commons	Fluoro	mUs][fAs][fG][fC][fA][mC][fA][mA]
		Pattern	[mU][fG][mG][mU][mA][fC][mG][mG]
		Modified	[mU][mU][mG][mCs][mGs][mG]
		Anti-sense
		Strand
KHK-860-	Hs-Mf-	Low-2′-	[MePhosphonate-4O-	822
898-729-	Mm-Rn	Fluoro	mUs][fAs][fGs][fC][fA][mG][fA][mC]
571	commons	Pattern	[mA][fC][mA][mU][mC][fU][mG][mG]
		Modified	[mC][mA][mG][mGs][mGs][mG]
		Anti-sense
		Strand
KHK-861-	Hs-Mf-	Low-2′-	[MePhosphonate-4O-	823
899-730-	Mm-Rn	Fluoro	mUs][fUs][fAs][fG][fC][mA][fG][mA]
572	commons	Pattern	[mC][fA][mC][mA][mU][fC][mU][mG]
		Modified	[mG][mC][mA][mGs][mGs][mG]
		Anti-sense
		Strand
KHK-865	Hs	Low-2′-	[MePhosphonate-4O-	824
	unique	Fluoro	mUs][fUs][fCs][fU][fG][mU][fA][mG]
		Pattern	[mC][fA][mG][mA][mC][fA][mC][mA]
		Modified	[mU][mC][mU][mGs][mGs][mG]
		Anti-sense
		Strand
KHK-882-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	825
920	commons	Fluoro	mUs][fGs][fAs][fU][fC][mA][fA][mC]
		Pattern	[mC][fU][mU][mC][mU][fC][mA][mA]
		Modified	[mA][mG][mU][mCs][mGs][mG]
		Anti-sense
		Strand
KHK-883-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	826
921	commons	Fluoro	mUs][fAs][fGs][fA][fU][mC][fA][mA]
		Pattern	[mC][fC][mU][mU][mC][fU][mC][mA]
		Modified	[mA][mA][mG][mUs][mGs][mG]
		Anti-sense
		Strand
KHK-885-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	827
923	commons	Fluoro	mUs][fUs][fCs][fA][fG][mA][fU][mC]
		Pattern	[mA][fA][mC][mC][mU][fU][mC][mU]
		Modified	[mC][mA][mA][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-1054-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	828
1092	commons	Fluoro	mUs][fUs][mCs][mU][fC][mC][fG][fU]
		Pattern	[mA][fG][mC][fC][mA][fA][mA][fC]
		Modified	[mA][mG][fC][mUs][mGs][mG]
		Anti-sense
		Strand
KHK-1075-	Hs-Mf-	Low-2′-	[MePhosphonate-4O-	829
1113-944-	Mm-Rn	Fluoro	mUs][fUs][fCs][fU][fU][mU][fG][mC]
786	commons	Pattern	[mU][fG][mA][mC][mA][fA][mA][mC]
		Modified	[mA][mC][mC][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-1078-	Hs-Mf-	Low-2′-	[MePhosphonate-4O-	830
1116-947-	Mm-Rn	Fluoro	mUs][fAs][fC][fA][fU][mC][fU][mU]
789	commons	Pattern	[mU][fG][mC][mU][mG][fA][mC][mA]
		Modified	[mA][mA][mC][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-1281-	Hs-Mf-	Low-2′-	[MePhosphonate-4O-	831
1319-1150-	Mm-Rn	Fluoro	mUs][fAs][fGs][fG][fC][mA][fU][mU]
992	commons	Pattern	[mG][fA][mA][mG][mG][fU][mG][mU]
		Modified	[mC][mU][mC][mCs][mGs][mG]
		Anti-sense
		Strand
KHK-1288-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	832
1326	commons	Fluoro	mUs][fAs][fUs][fG][fA][mC][fG][mG]
		Pattern	[mA][fG][mG][mC][mA][fU][mU][mG]
		Modified	[mA][mA][mG][mGs][mGs][mG]
		Anti-sense
		Strand
KHK-1290-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	833
1328	commons	Fluoro	mUs][fAs][fGs][fA][fU][mG][fA][mC]
		Pattern	[mG][fG][mA][mG][mG][fC][mA][mU]
		Modified	[mU][mG][mA][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-1148-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	834
1186	commons	Fluoro	mUs][fCs][fCs][fC][fU][mU][fU][mC]
		Pattern	[mC][fU][mC][mA][mC][fA][mC][mG]
		Modified	[mA][mC][mC][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-1152-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	835
1190	commons	Fluoro	mUs][fCs][fAs][fG][fC][mC][fC][mC]
		Pattern	[mU][fU][mU][mC][mC][fU][mC][mA]
		Modified	[mC][mA][mC][mGs][mGs][mG]
		Anti-sense
		Strand
KHK-1154-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	836
1192	commons	Fluoro	mUs][fCs][fAs][fC][fA][mG][fC][mC]
		Pattern	[mC][fC][mU][mU][mU][fC][mC][mU]
		Modified	[mC][mA][mC][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-1155-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	837
1193	commons	Fluoro	mUs][fGs][fCs][fA][fC][mA][fG][mC]
		Pattern	[mC][fC][mC][mU][mU][fU][mC][mC]
		Modified	[mU][mC][mA][mCs][mGs][mG]
		Anti-sense
		Strand
KHK-1277	Hs	Low-2′-	[MePhosphonate-4O-	838
	unique	Fluoro	mUs][fAs][fUs][fU][fG][mA][fA][mG]
		Pattern	[mG][fU][mG][mU][mC][fU][mC][mC]
		Modified	[mA][mG][mC][mUs][mGs][mG]
		Anti-sense
		Strand
KHK-1147-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	839
1185	commons	Fluoro	mUs][fCs][fCs][fU][fU][mU][fC][mC]
		Pattern	[mU][fC][mA][mC][mA][fC][mG][mA]
		Modified	[mC][mC][mA][mUs][mGs][mG]
		Anti-sense
		Strand
KHK-869-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	840
934	commons	Fluoro	mUs][fAs][fAs][fA][fG][mU][fC][mU]
		Pattern	[mG][fU][mA][mG][mC][fA][mG][mA]
		Modified	[mC][mA][mC][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-873	Hs	Low-2′-	[MePhosphonate-4O-	841
	unique	Fluoro	mUs][fUs][fCs][fU][fC][mA][fA][mA]
		Pattern	[mG][fU][mC][mU][mG][fU][mA][mG]
		Modified	[mC][mA][mG][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-879	Hs	Low-2′-	[MePhosphonate-4O-	842
	unique	Fluoro	mUs][fCs][fAs][fA][fC][mC][fU][mU]
		Pattern	[mC][fU][mC][mA][mA][fA][mG][mU]
		Modified	[mC][mU][mG][mUs][mGs][mG]
		Anti-sense
		Strand
KHK-881	Hs	Low-2′-	[MePhosphonate-4O-	843
	unique	Fluoro	mUs][fAs][fUs][fC][fA][mA][fC][mC]
		Pattern	[mU][fU][mC][mU][mC][fA][mA][mA]
		Modified	[mG][mU][mC][mUs][mGs][mG]
		Anti-sense
		Strand
KHK-896-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	844
934	commons	Fluoro	mUs][fCs][fUs][fU][fG][mA][fA][mC]
		Pattern	[mU][fG][mG][mG][mU][fC][mA][mG]
		Modified	[mA][mU][mC][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-1064-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	845
1102	commons	Fluoro	mUs][fAs][fAs][fA][fC][mA][fC][mC]
		Pattern	[mA][fC][mG][mU][mC][fU][mC][mC]
		Modified	[mG][mU][mA][mGs][mGs][mG]
		Anti-sense
		Strand
KHK-1077-	Hs-Mf-	Low-2′-	[MePhosphonate-4O-	846
1115-946-	Mm-Rn	Fluoro	mUs][fCs][fAs][fU][fC][mU][fU][mU]
788	commons	Pattern	[mG][fC][mU][mG][mA][fC][mA][mA]
		Modified	[mA][mC][mA][mCs][mGs][mG]
		Anti-sense
		Strand
KHK-1080-	Hs-Mf-	Low-2′-	[MePhosphonate-4O-	847
1118-949-	Mm-Rn	Fluoro	mUs][fCs][fCs][fA][fC][mA][fU][mC]
791	commons	Pattern	[mU][fU][mU][mG][mC][fU][mG][mA]
		Modified	[mC][mA][mA][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-1106-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	848
1144	commons	Fluoro	mUs][fUs][fGs][fC][fU][mG][fA][mC]
		Pattern	[mU][fG][mG][mA][mA][fC][mC][mC]
		Modified	[mC][mA][mA][mGs][mGs][mG]
		Anti-sense
		Strand
KHK-1334-	Hs-Mf	Low-2′-	[MePhosphonate-4O-	849
1372	commons	Fluoro	mUs][fGs][fAs][fA][fU][mC][fU][mC]
		Pattern	[mA][fG][mU][mG][mC][fU][mU][mC]
		Modified	[mC][mU][mG][mCs][mGs][mG]
		Anti-sense
		Strand
KHK-516-	Hs-Mf-	Med-2′-	[MePhosphonate-4O-	850
554-385-	Mm-Rn	Fluoro	mUs][fAs][fCs][fA][fG][mG][fA][fU]
227	commons	Pattern	[mC][fU][mG][mC][mU][fU][mC][fU]
		Modified	[mC][mU][fU][mCs][mGs][mG]
		Anti-sense
		Strand
KHK-804	Hs	Med-2′-	[MePhosphonate-4O-	851
	unique	Fluoro	mUs][fAs][fGs][fU][fU][mG][fU][fU]
		Pattern	[mG][fA][mU][mG][mA][fU][mG][fC]
		Modified	[mA][mG][fC][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-829-	Hs-Mf	Med-2′-	[MePhosphonate-4O-	852
838	commons	Fluoro	mUs][fAs][fGs][fC][fA][mC][fA][fA]
		Pattern	[mU][fG][mG][mU][mA][fC][mG][fG]
		Modified	[mU][mU][fG][mCs][mGs][mG]
		Anti-sense
		Strand
KHK-861-	Hs-Mf-	Med-2′-	[MePhosphonate-4O-	853
899-730-	Mm-Rn	Fluoro	mUs][fUs][fAs][fG][fC][mA][fG][fA]
572	commons	Pattern	[mC][fA][mC][mA][mU][fC][mU][fG]
		Modified	[mG][mC][fA][mGs][mGs][mG]
		Anti-sense
		Strand
KHK-865	Hs	Med-2′-	[MePhosphonate-4O-	854
	unique	Fluoro	mUs][fUs][fCs][fU][fG][mU][fA][fG]
		Pattern	[mC][fA][mG][mA][mC][fA][mC][fA]
		Modified	[mU][mC][fU][mGs][mGs][mG]
		Anti-sense
		Strand
KHK-882-	Hs-Mf	Med-2′-	[MePhosphonate-4O-	855
920	commons	Fluoro	mUs][fGs][fAs][fU][fC][mA][fA][fC]
		Pattern	[mC][fU][mU][mC][mU][fC][mA][fA]
		Modified	[mA][mG][fU][mCs][mGs][mG]
		Anti-sense
		Strand
KHK-883-	Hs-Mf	Med-2′-	[MePhosphonate-4O-	856
921	commons	Fluoro	mUs][fAs][fGs][fA][fU][mC][fA][fA]
		Pattern	[mC][fC][mU][mU][mC][fU][mC][fA]
		Modified	[mA][mA][fG][mUs][mGs][mG]
		Anti-sense
		Strand
KHK-885-	Hs-Mf	Med-2′-	[MePhosphonate-4O-	857
923	commons	Fluoro	mUs][fUs][fCs][fA][fG][mA][fU][fC]
		Pattern	[mA][fA][mC][mC][mU][fU][mC][fU]
		Modified	[mC][mA][fA][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-1054-	Hs-Mf	Med-2′-	[MePhosphonate-4O-	858
1092	commons	Fluoro	mUs][fUs][mCs][mU][fC][mC][fG][fU]
		Pattern	[mA][fG][mC][fC][mA][fA][mA][fC]
		Modified	[mA][mG][fC][mUs][mGs][mG]
		Anti-sense
		Strand
KHK-1076-	Hs-Mf-	Med-2′-	[MePhosphonate-4O-	859
1114-945-	Mm-Rn	Fluoro	mUs][fAs][fUs][fC][fU][mU][fU][fG]
787	commons	Pattern	[mC][fU][mG][mA][mC][fA][mA][fA]
		Modified	[mC][mA][fC][mCs][mGs][mG]
		Anti-sense
		Strand
KHK-1078-	Hs-Mf-	Med-2′-	[MePhosphonate-4O-	860
1116-947-	Mm-Rn	Fluoro	mUs][fAs][fCs][fA][fU][mC][fU][fU]
789	commons	Pattern	[mU][fG][mC][mU][mG][fA][mC][fA]
		Modified	[mA][mA][fC][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-1288-	Hs-Mf	Med-2′-	[MePhosphonate-4O-	861
1326	commons	Fluoro	mUs][fAs][fUs][fG][fA][mC][fG][fG]
		Pattern	[mA][fG][mG][mC][mA][fU][mU][fG]
		Modified	[mA][mA][fG][mGs][mGs][mG]
		Anti-sense
		Strand
KHK-1290-	Hs-Mf	Med-2′-	[MePhosphonate-4O-	862
1328	commons	Fluoro	mUs][fAs][fGs][fA][fU][mG][fA][fC]
		Pattern	[mG][fG][mA][mG][mG][fC][mA][fU]
		Modified	[mU][mG][fA][mAs][mGs][mG]
		Anti-sense
		Strand
KHK-1334-	Hs-Mf	Med-2′-	[MePhosphonate-4O-	863
1372	commons	Fluoro	mUs][fGs][fAs][fA][fU][mC][fU][fC]
		Pattern	[mA][fG][mU][mG][mC][fU][mU][fC]
		Modified	[mC][mU][fG][mCs][mGs][mG]
		Anti-sense
		Strand
Fam Probe	Fam	N/A	CCCTGGCCATGTTG	864
	Probe
	for NHP
	KHK
	primers
Forward-	Mouse	N/A	TGGAGGTGGAGAAGCCA	865
1026	KHK
Primer
Reverse-	Mouse	N/A	GACCATACAAGCCCCTCAAG	866
1157	KHK
Primer
Probe-	Probe	N/A	TGGTGTTTGTCAGCAAAGATGTGGC	867
1080	for
	Mouse
	KHK
	primers
Forward-	5′	N/A	AGGAAGCTCTGGGAGTA	868
496	Assay
Primer
Reverse-	5′	N/A	CCTCCTTAGGGTACTTGTC	869
596	Assay
Primer
Probe-518	5′	N/A	ATGGAAGAGAAGCAGATCCTGTGCG	870
	Assay
	Probe
Stem Loop	Un-	N/A	GCAGCCGAAAGGCUGC	871
	modified
	stem
	loop
	sequence
KHK-1277	Hs	36mer	AGCUGGAGACACCUUCAAUAGCAGCCGA	872
	unique	Sense	AAGGCUGC
		Strand
KHK-1155-	Hs-Mf	36mer	GUGAGGAAAGGGGCUGUGCAGCAGCCGA	873
1193	commons	Sense	AAGGCUGC
		Strand
KHK-1152-	Hs-Mf	36mer	CGUGUGAGGAAAGGGGCUGAGCAGCCGA	874
1190	commons	Sense	AAGGCUGC
		Strand
KHK-881	Hs	36mer	AGACUUUGAGAAGGUUGAUAGCAGCCGA	875
	unique	Sense	AAGGCUGC
		Strand
KHK-879	Hs	36mer	ACAGACUUUGAGAAGGUUGAGCAGCCGA	876
	unique	Sense	AAGGCUGC
		Strand
KHK-869	Hs-Mf	36mer	UGUGUCUGCUACAGACUUUAGCAGCCGA	877
	commons	Sense	AAGGCUGC
		Strand
KHK-873	Hs	36mer	UCUGCUACAGACUUUGAGAAGCAGCCGA	878
	unique	Sense	AAGGCUGC
		strand
KHK-1277	Hs	22mer	UAUUGAAGGUGUCUCCAGCUGG	879
	unique	anti-sense
		strand
KHK-1155-	Hs-Mf	22mer	UGCACAGCCCCUUUCCUCACGG	880
1193	commons	anti-sense
		strand
KHK-1152-	Hs-Mf	22mer	UCAGCCCCUUUCCUCACACGGG	881
1190	commons	anti-sense
		strand
KHK-881	Hs	22mer	UAUCAACCUUCUCAAAGUCUGG	882
	unique	anti-sense
		strand
KHK-879	Hs	22mer	UCAACCUUCUCAAAGUCUGUGG	883
	unique	anti-sense
		strand
KHK-869	Hs-Mf	22mer	UAAAGUCUGUAGCAGACACAGG	884
	commons	anti-sense
		strand
KHK-873	Hs	22mer	UUCUCAAAGUCUGUAGCAGAGG	885
	unique	anti-sense
		strand
KHK-510-	Hs-Mf-	36mer	CUCAUGGAAGAGAAGCAGAAGCAGCCGA	886
548-379-	Mm-Rn	Sense	AAGGCUGC
221	commons	strand
KHK-516-	Hs-Mf-	36mer	GAAGAGAAGCAGAUCCUGUAGCAGCCGA	887
554-385-	Mm-Rn	Sense	AAGGCUGC
227	commons	strand
KHK-829-	Hs-Mf	36mer	GCAACCGUACCAUUGUGCUAGCAGCCGA	888
838	commons	Sense	AAGGCUGC
		strand
KHK-860-	Hs-Mf-	36mer	CCUGCCAGAUGUGUCUGCUAGCAGCCGA	889
898-729-	Mm-Rn	Sense	AAGGCUGC
571	commons	strand
KHK-861-	Hs-Mf-	36mer	CUGCCAGAUGUGUCUGCUAAGCAGCCGA	890
899-730-	Mm-Rn	Sense	AAGGCUGC
572	commons	strand
KHK-865	Hs	36mer	CAGAUGUGUCUGCUACAGAAGCAGCCGA	891
	unique	Sense	AAGGCUGC
		strand
KHK-882-	Hs-Mf	36mer	GACUUUGAGAAGGUUGAUCAGCAGCCGA	892
920	commons	Sense	AAGGCUGC
		strand
KHK-883-	Hs-Mf	36mer	ACUUUGAGAAGGUUGAUCUAGCAGCCGA	893
921	commons	Sense	AAGGCUGC
		strand
KHK-885-	Hs-Mf	36mer	UUUGAGAAGGUUGAUCUGAAGCAGCCGA	894
923	commons	Sense	AAGGCUGC
		strand
KHK-1054-	Hs-Mf	36mer	AGCUGUUUGGCUACGGAGAAGCAGCCGA	895
1092	commons	Sense	AAGGCUGC
		strand
KHK-1075-	Hs-Mf-	36mer	UGGUGUUUGUCAGCAAAGAAGCAGCCGA	896
1113-944-	Mm-Rn	Sense	AAGGCUGC
786	commons	strand
KHK-1078-	Hs-Mf-	36mer	UGUUUGUCAGCAAAGAUGUAGCAGCCGA	897
1116-947-	Mm-Rn	Sense	AAGGCUGC
789	commons	strand
KHK-1281-	Hs-Mf-	36mer	GGAGACACCUUCAAUGCCUAGCAGCCGA	898
1319-1150-	Mm-Rn	Sense	AAGGCUGC
992	commons	strand
KHK-1288-	Hs-Mf	36mer	CCUUCAAUGCCUCCGUCAUAGCAGCCGA	899
1326	commons	Sense	AAGGCUGC
		strand
KHK-1290-	Hs-Mf	36mer	UUCAAUGCCUCCGUCAUCUAGCAGCCGA	900
1328	commons	Sense	AAGGCUGC
		strand
KHK-1148-	Hs-Mf	36mer	UGGUCGUGUGAGGAAAGGGAGCAGCCGA	901
1186	commons	Sense	AAGGCUGC
		strand
KHK-1154-	Hs-Mf	36mer	UGUGAGGAAAGGGGCUGUGAGCAGCCGA	902
1192	commons	Sense	AAGGCUGC
		strand
KHK-1147-	Hs-Mf	36mer	AUGGUCGUGUGAGGAAAGGAGCAGCCGA	903
1185	commons	Sense	AAGGCUGC
		strand
KHK-896-	Hs-Mf	36mer	UGAUCUGACCCAGUUCAAGAGCAGCCGA	904
934	commons	Sense	AAGGCUGC
		strand
KHK-1064-	Hs-Mf	36mer	CUACGGAGACGUGGUGUUUAGCAGCCGA	905
1102	commons	Sense	AAGGCUGC
		strand
KHK-1077-	Hs-Mf-	36mer	GUGUUUGUCAGCAAAGAUGAGCAGCCGA	906
1115-946-	Mm-Rn	Sense	AAGGCUGC
788	commons	strand
KHK-1080-	Hs-Mf-	36mer	UUUGUCAGCAAAGAUGUGGAGCAGCCGA	907
11 18-949-	Mm-Rn	Sense	AAGGCUGC
791	commons	strand
KHK-1106-	Hs-Mf	36mer	CUUGGGGUUCCAGUCAGCAAGCAGCCGA	908
1144	commons	Sense	AAGGCUGC
		strand
KHK-1334-	Hs-Mf	36mer	GCAGGAAGCACUGAGAUUCAGCAGCCGA	909
1372	commons	Sense	AAGGCUGC
		strand
KHK-804	Hs	36mer	UGCUGCAUCAUCAACAACUAGCAGCCGAA	910
	unique	Sense	AGGCUGC
		strand
KHK-1076-	Hs-Mf-	36mer	GGUGUUUGUCAGCAAAGAUAGCAGCCGA	911
1114-945-	Mm-Rn	Sense	AAGGCUGC
787	commons	strand
KHK-510-	Hs-Mf-	22mer	UUCUGCUUCUCUUCCAUGAGGG	912
548-379-	Mm-Rn	anti-sense
221	commons	strand
KHK-516-	Hs-Mf-	22mer	UACAGGAUCUGCUUCUCUUCGG	913
554-385-	Mm-Rn	anti-sense
227	commons	strand
KHK-829-	Hs-Mf	22mer	UAGCACAAUGGUACGGUUGCGG	914
838	commons	anti-sense
		strand
KHK-860-	Hs-Mf-	22mer	UAGCAGACACAUCUGGCAGGGG	915
898-729-	Mm-Rn	anti-sense
571	commons	strand
KHK-861-	Hs-Mf-	22mer	UUAGCAGACACAUCUGGCAGGG	916
899-730-	Mm-Rn	anti-sense
572	commons	strand
KHK-865	Hs	22mer	UUCUGUAGCAGACACAUCUGGG	917
	unique	anti-sense
		strand
KHK-882-	Hs-Mf	22mer	UGAUCAACCUUCUCAAAGUCGG	918
920	commons	anti-sense
		strand
KHK-883-	Hs-Mf	22mer	UAGAUCAACCUUCUCAAAGUGG	919
921	commons	anti-sense
		strand
KHK-885-	Hs-Mf	22mer	UUCAGAUCAACCUUCUCAAAGG	920
923	commons	anti-sense
		strand
KHK-1054-	Hs-Mf	22mer	UUCUCCGUAGCCAAACAGCUGG	921
1092	commons	anti-sense
		strand
KHK-1075-	Hs-Mf-	22mer	UUCUUUGCUGACAAACACCAGG	922
1113-944-	Mm-Rn	anti-sense
786	commons	strand
KHK-1078-	Hs-Mf-	22mer	UACAUCUUUGCUGACAAACAGG	923
1116-947-	Mm-Rn	anti-sense
789	commons	strand
KHK-1281-	Hs-Mf-	22mer	UAGGCAUUGAAGGUGUCUCCGG	924
1319-1150-	Mm-Rn	anti-sense
992	commons	strand
KHK-1288-	Hs-Mf	22mer	UAUGACGGAGGCAUUGAAGGGG	925
1326	commons	anti-sense
		strand
KHK-1290-	Hs-Mf	22mer	UAGAUGACGGAGGCAUUGAAGG	926
1328	commons	anti-sense
		strand
KHK-1148-	Hs-Mf	22mer	UCCCUUUCCUCACACGACCAGG	927
1186	commons	anti-sense
		strand
KHK-1154-	Hs-Mf	22mer	UCACAGCCCCUUUCCUCACAGG	928
1192	commons	anti-sense
		strand
KHK-1147-	Hs-Mf	22mer	UCCUUUCCUCACACGACCAUGG	929
1185	commons	anti-sense
		strand
KHK-873	Hs	22mer	UUCUCAAAGUCUGUAGCAGAGG	930
	unique	anti-sense
		strand
KHK-896-	Hs-Mf	22mer	UGUUGAACUGGGUCAGAUCAGG	931
934	commons	anti-sense
		strand
KHK-1064-	Hs-Mf	22mer	UAAACACCACGUCUCCGUAGGG	932
1102	commons	anti-sense
		strand
KHK-1077-	Hs-Mf-	22mer	UCAUCUUUGCUGACAAACACGG	933
1115-946-	Mm-Rn	anti-sense
788	commons	strand
KHK-1080-	Hs-Mf-	22mer	UCCACAUCUUUGCUGACAAAGG	934
1118-949-	Mm-Rn	anti-sense
791	commons	strand
KHK-1106-	Hs-Mf	22mer	UUGCUGACUGGAACCCCAAGGG	935
1144	commons	anti-sense
		strand
KHK-1334-	Hs-Mf	22mer	UGAAUCUCAGUGCUUCCUGCGG	936
1372	commons	anti-sense
		strand
KHK-804	Hs	22mer	UAGUUGUUGAUGAUGCAGCAGG	937
	unique	anti-sense
		strand
KHK-1076-	Hs-Mf-	22mer	UAUCUUUGCUGACAAACACCGG	938
1114-945-	Mm-Rn	anti-sense
787	commons	strand
MmKHK-	Forward	N/A	GCTCTTCCAGTTGTTTAGCTATGGT	939
ALL-5-6
MmKHK-	Reverse	N/A	CAGGTGCTTGGCCACATCTT	940
ALL-5-6
MmKHK-	Probe	N/A	AGGTGGTGTTTGTCAGC	941
ALL-5-6
KHK-516-	Hs-Mf-	19mer	GAAGAGAAGCAGAUCCUGU	942
554-385-	Mm-Rn	Sense
227	commons	strand
KHK-865	Hs	19mer	CAGAUGUGUCUGCUACAGA	943
	unique	Sense
		strand
KHK-882-	Hs-Mf	19mer	GACUUUGAGAAGGUUGAUC	944
920	commons	Sense
		strand
KHK-885-	Hs-Mf	19mer	UUUGAGAAGGUUGAUCUGA	945
923	commons	Sense
		strand
KHK-1078-	Hs-Mf-	19mer	UGUUUGUCAGCAAAGAUGU	946
1116-947-	Mm-Rn	Sense
789	commons	strand
KHK-1334-	Hs-Mf	19mer	GCAGGAAGCACUGAGAUUC	947
1372	commons	Sense
		strand
KHK-516-	Hs-Mf-	19mer	ACAGGAUCUGCUUCUCUUC	948
554-385-	Mm-Rn	anti-sense
227	commons	strand
KHK-865	Hs	19mer	UCUGUAGCAGACACAUCUG	949
	unique	anti-sense
		strand
KHK-882-	Hs-Mf	19mer	GAUCAACCUUCUCAAAGUC	950
920	commons	anti-sense
		strand
KHK-885-	Hs-Mf	19mer	UCAGAUCAACCUUCUCAAA	951
923	commons	anti-sense
		strand
KHK-1078-	Hs-Mf-	19mer	ACAUCUUUGCUGACAAACA	952
1116-947-	Mm-Rn	anti-sense
789	commons	strand
KHK-1334-	Hs-Mf	19mer	GAAUCUCAGUGCUUCCUGC	953
1372	commons	anti-sense
		strand

Particular Aspects and Embodiments of the Present Invention are Described with Reference to the Following Clauses:1. A double stranded RNAi oligonucleotide for reducing ketohexokinase (KHK) expression, the oligonucleotide comprising an antisense strand and a sense strand, wherein the antisense strand and the sense strand form a duplex region, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387 and wherein the region of complementarity is at least 15 contiguous nucleotides in length, or a pharmaceutically acceptable salt thereof.2. The RNAi oligonucleotide of clause 1, wherein the sense strand comprises a sequence set forth in any one of SEQ ID NOs: 4-387.3. The RNAi oligonucleotide of clause 1 or 2, wherein the antisense strand comprises a sequence set forth in any one of SEQ ID NOs: 388-771.4. A double stranded RNAi oligonucleotide for inhibiting expression of KHK, wherein said double stranded RNAi oligonucleotide comprises a sense strand and an antisense strand forming a duplex region, wherein said sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the nucleotide sequences of SEQ ID NO:4-387 and said antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the nucleotide sequences of SEQ ID NO: 388-771, or a pharmaceutically acceptable salt thereof.5. The RNAi oligonucleotide of any one of clauses 1-4, wherein the sense strand is 15 to 50 nucleotides in length.6. The RNAi oligonucleotide of any one of clauses 1-4, wherein the sense strand is 18 to 36 nucleotides in length.7. The RNAi oligonucleotide of any one of clauses 1-4, wherein the sense strand is 15 to 30 nucleotides in length.8. The RNAi oligonucleotide of any one of clauses 1-7, wherein the antisense strand is 15-30 nucleotides in length.9. The RNAi oligonucleotide of any one of clauses 1-8, wherein the antisense strand and the sense strand form a duplex region of at least 19 nucleotides in length, optionally at least 20 nucleotides in length.10. The RNAi oligonucleotide of any one of clauses 1-3 and 5-9, wherein the region of complementarity is at least 19 contiguous nucleotides in length, optionally at least 20 nucleotides in length.11. A double stranded RNAi oligonucleotide for reducing KHK expression, the oligonucleotide comprising:

• • (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is selected from SEQ ID NOs: 948-953, and
  • (ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand, wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.12. The RNAi oligonucleotide of any one of clauses 1-11, wherein the sense strand comprises at its 3′ end a stem-loop set forth as: S1-L-S2, wherein S1 is complementary to

S2, and wherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length.

13. The RNAi oligonucleotide of clause 12, wherein L is a triloop or a tetraloop.14. The RNAi oligonucleotide of clause 13, wherein L is a tetraloop.15. The RNAi oligonucleotide of clause 14, wherein the tetraloop comprises the sequence 5′-GAAA-3′.16. The RNAi oligonucleotide of any one of clauses 12-15, wherein the S1 and S2 are 1-10 nucleotides in length and have the same length.17. The RNAi oligonucleotide of clause 16, wherein S1 and S2 are 1 nucleotide, 2 nucleotides, 3 nucleotides, 4 nucleotides, 5 nucleotides, 6 nucleotides, 7 nucleotides, 8 nucleotides, 9 nucleotides, or 10 nucleotides in length.18. The RNAi oligonucleotide of clause 17, wherein S1 and S2 are 6 nucleotides in length.19. The RNAi oligonucleotide of any one of clauses 12-18, wherein the stem-loop comprises the sequence 5′-GCAGCCGAAAGGCUGC-3′ (SEQ ID NO: 871).20. The RNAi oligonucleotide of any one of clauses 1-19, comprising a nicked tetraloop structure.21. The RNAi oligonucleotide of any one of clauses 1-19, comprising a nick between the 3′ terminus of the sense strand and the 5′ terminus of the antisense strand.22. The RNAi oligonucleotide of any one of clauses 1-21, wherein the antisense and sense strands are not covalently linked.23. The RNAi oligonucleotide of any one of clauses 1-10 and 12-22, wherein the antisense strand comprises an overhang of one or more nucleotides in length at the 3′ terminus.24. The RNAi oligonucleotide of any one of clauses 11-23, wherein the overhang comprises purine nucleotides.25. The RNAi oligonucleotide of any one of clauses 11-24, wherein the overhang is 2 nucleotides in length.26. The RNAi oligonucleotide of clause 25, wherein the 3′ overhang is selected from AA, GG, AG, and GA.27. The RNAi oligonucleotide of clause 26, wherein the overhang is GG or AA.28. The RNAi oligonucleotide of clause 26, wherein the overhang is GG.29. The RNAi oligonucleotide of any one of the preceding clauses, wherein the oligonucleotide comprises at least one modified nucleotide.30. The RNAi oligonucleotide of clause 29, wherein the modified nucleotide comprises a 2′-modification.31. The RNAi oligonucleotide of clause 30, wherein the 2′-modification is a modification selected from 2′-aminoethyl, 2′-fluoro, 2′-O-methyl, 2′-O-methoxyethyl, and 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid.32. The RNAi oligonucleotide of any one of clauses 29-31, wherein about 10-15%, 10%, 11%, 12%, 13%, 14% or 15% of the nucleotides of the sense strand comprise a 2′-fluoro modification.33. The RNAi oligonucleotide of any one of clauses 29-32, wherein about 25-35%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34% or 35% of the nucleotides of the antisense strand comprise a 2′-fluoro modification.34. The RNAi oligonucleotide of any one of clauses 29-33, wherein about 25-35%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34% or 35% of the nucleotides of the oligonucleotide comprise a 2′-fluoro modification.35. The RNAi oligonucleotide of any one of clauses 29-34, wherein all the nucleotides of the oligonucleotide are modified.36. The RNAi oligonucleotide of any one of clauses 29-34, wherein the sense strand comprises 36 nucleotides with positions 1-36 numbered from 5′ to 3′, wherein positions 8, 9, 10 and 11 of the sense strand are modified.37. The RNAi oligonucleotide of any one of clauses 29-34, wherein the sense strand comprises 36 nucleotides with positions 1-36 numbered from 5′ to 3′, wherein positions 3, 8, 9, 10, 12, 13 and 17 of the sense strand are modified.38. The RNAi oligonucleotide of any one of clauses 29-34, wherein the antisense strand comprises 22 nucleotides with positions 1-22 numbered from 5′ to 3′, and wherein positions 2, 3, 4, 5, 7, 10 and 14 of the antisense strand are modified.39. The RNAi oligonucleotide of any one of clauses 29-34, wherein the antisense strand comprises 22 nucleotides with positions 1-22 numbered from 5′ to 3′, and wherein positions 2-5, 7, 8, 10, 14, 16 and 19 of the antisense strand are modified.40. The RNAi oligonucleotide of any one of clauses 36-39, where the modification is 2′-fluoro.41. The RNAi oligonucleotide of any one of clauses 32-34 and 36-40, wherein the remaining nucleotides comprise a 2′-O-methyl modification.42. The RNAi oligonucleotide of any one of the preceding clauses, wherein the oligonucleotide comprises at least one modified internucleotide linkage.43. The RNAi oligonucleotide of clause 42, wherein the at least one modified internucleotide linkage is a phosphorothioate linkage.44. The RNAi oligonucleotide of clause 43, wherein the antisense strand comprises a phosphorothioate linkage (i) between positions 1 and 2, and between positions 2 and 3; or (ii) between positions 1 and 2, between positions 2 and 3, and between positions 3 and 4, wherein positions are numbered 1-4 from 5′ to 3′.45. The RNAi oligonucleotide of clause 43 or 44, wherein the antisense strand is 22 nucleotides in length, and wherein the antisense strand comprises a phosphorothioate linkage between positions 20 and 21 and between positions 21 and 22, wherein positions are numbered 1-22 from 5′ to 3′.46. The RNAi oligonucleotide of any one of clauses 1-45, wherein the antisense strand comprises a phosphorylated nucleotide at the 5′ terminus, wherein the phosphorylated nucleotide is selected from uridine and adenosine.47. The RNAi oligonucleotide of clause 46, wherein the phosphorylated nucleotide is uridine.48. The RNAi oligonucleotide of any one of the preceding clauses, wherein the 4′-carbon of the sugar of the 5′-terminal nucleotide of the antisense strand comprises a phosphate analog.49. The RNAi oligonucleotide of clause 48, wherein the phosphate analog is oxymethylphosphonate, vinylphosphonate or malonyl phosphonate, optionally wherein the phosphate analog is a 4′-phosphate analog comprising 5′-methoxyphosphonate-4′-oxy.50. The RNAi oligonucleotide of any one of the preceding clauses, wherein at least one nucleotide of the oligonucleotide is conjugated to one or more targeting ligands.51. The RNAi oligonucleotide of clause 50, wherein each targeting ligand comprises a carbohydrate, amino sugar, cholesterol, polypeptide, or lipid.52. The RNAi oligonucleotide of any one of clauses 11-51, wherein the stem loop comprises one or more targeting ligands conjugated to one or more nucleotides of the stem loop.53. The RNAi oligonucleotide of clause 52, wherein the one or more targeting ligands is conjugated to one or more nucleotides of the loop.54. The RNAi oligonucleotide of clause 53, wherein the loop comprises 4 nucleotides numbered 1-4 from 5′ to 3′, wherein nucleotides at positions 2, 3, and 4 each comprise one or more targeting ligands, wherein the targeting ligands are the same or different.55. The RNAi oligonucleotide of any one of clauses 50-54, wherein each targeting ligand comprises a N-acetylgalactosamine (GalNAc) moiety.56. The RNAi oligonucleotide of clause 55, wherein the GalNAc moiety is a monovalent GalNAc moiety, a bivalent GalNAc moiety, a trivalent GalNAc moiety or a tetravalent GalNAc moiety.57. The RNAi oligonucleotide of any one of clauses 11-56, wherein up to 4 nucleotides of L of the stem-loop are each conjugated to a monovalent GalNAc moiety.58. The RNAi oligonucleotide of any one of clauses 1-57, wherein the region of complementarity comprised by the antisense strand is fully complementary to the KHK mRNA target sequence at nucleotide positions 2-8 of the antisense strand, wherein nucleotide positions are numbered 5′ to 3′.59. The RNAi oligonucleotide of any one of clauses 1-57, wherein the region of complementarity comprised by the antisense strand is fully complementary to the KHK mRNA target sequence at nucleotide positions 2-11 of the antisense strand, wherein nucleotide positions are numbered 5′ to 3′.60. The RNAi oligonucleotide of any one of clauses 1-59, wherein the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs: 872-878 and 886-911.61. The RNAi oligonucleotide of any one of clauses 1-60, wherein the antisense strand comprises a nucleotide sequence of any one of SEQ ID NOs: 879-884 and 912-938.62. The RNAi oligonucleotide of any one of clauses 1-61, wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 886 and 912, respectively;
  • (b) SEQ ID NOs: 887 and 913, respectively;
  • (c) SEQ ID NOs: 910 and 937, respectively;
  • (d) SEQ ID NOs: 888 and 914, respectively;
  • (e) SEQ ID NOs: 889 and 915, respectively;
  • (f) SEQ ID NOs: 890 and 916, respectively;
  • (g) SEQ ID NOs: 891 and 917, respectively;
  • (h) SEQ ID NOs: 877 and 884, respectively;
  • (i) SEQ ID NOs: 878 and 930, respectively;
  • (j) SEQ ID NOs: 876 and 883, respectively;
  • (k) SEQ ID NOs: 875 and 882, respectively;
  • (l) SEQ ID NOs: 892 and 918, respectively;
  • (m) SEQ ID NOs: 893 and 919, respectively;
  • (n) SEQ ID NOs: 894 and 920, respectively;
  • (o) SEQ ID NOs: 904 and 931, respectively;
  • (p) SEQ ID NOs: 895 and 921, respectively;
  • (q) SEQ ID NOs: 905 and 932, respectively;
  • (r) SEQ ID NOs: 896 and 922, respectively;
  • (s) SEQ ID NOs: 911 and 938, respectively;
  • (t) SEQ ID NOs: 906 and 933, respectively;
  • (u) SEQ ID NOs: 897 and 923, respectively;
  • (v) SEQ ID NOs: 907 and 934, respectively;
  • (w) SEQ ID NOs: 908 and 935, respectively;
  • (x) SEQ ID NOs: 903 and 929, respectively;
  • (y) SEQ ID NOs: 901 and 927, respectively;
  • (z) SEQ ID NOs: 874 and 881, respectively;
  • (aa) SEQ ID NOs: 902 and 928, respectively;
  • (bb) SEQ ID NOs: 873 and 880, respectively;
  • (cc) SEQ ID NOs: 872 and 879, respectively;
  • (dd) SEQ ID NOs: 898 and 924, respectively;
  • (ee) SEQ ID NOs: 899 and 925, respectively;
  • (ff) SEQ ID NOs: 900 and 926, respectively; and
  • (gg) SEQ ID NOs: 909 and 936, respectively.63. The RNAi oligonucleotide of any one of clauses 1-62, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 909 and 936, respectively.64. The RNAi oligonucleotide of any one of clauses 1-62, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 894 and 920, respectively.65. The RNAi oligonucleotide of any one of clauses 1-62, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 897 and 923, respectively.66. The RNAi oligonucleotide of any one of clauses 1-62, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 892 and 918, respectively.67. The RNAi oligonucleotide of any one of clauses 1-62, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 891 and 917, respectively.68. The RNAi oligonucleotide of any one of clauses 1-62, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 887 and 913, respectively.69. The RNAi oligonucleotide of any one of clauses 1-59, wherein the antisense strand is 22 nucleotides in length.70. The RNAi oligonucleotide of clause 69, wherein the antisense strand comprises a nucleotide sequence selected from SEQ ID NOs: 913, 917, 918, 920, 923 and 936.71. The RNAi oligonucleotide of any one of clauses 1-59 and 69-70, wherein the sense strand comprises a nucleotide sequence selected from SEQ ID NOs: 942-947.72. The RNAi oligonucleotide of any one of clauses 1-59 and 69-71, wherein the sense strand is 36 nucleotides in length.73. The RNAi oligonucleotide of clause 72, wherein the sense strand comprises a nucleotide sequence selected from SEQ ID NOs: 887, 891, 892, 894, 897 and 909.74. The RNAi oligonucleotide of any one of clauses 60-73, wherein the sense strand and the antisense strand are modified, wherein the antisense strand and the sense strand comprise one or more 2′-fluoro and 2′-O-methyl modified nucleotides and at least one phosphorothioate linkage, wherein the 4′-carbon of the sugar of the 5′-nucleotide of the antisense strand comprises a phosphate analog.75. The RNAi oligonucleotide of any one of clauses 1-59, wherein the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs: 774-804.76. The RNAi oligonucleotide of any one of clauses 1-59 and 75, wherein the antisense strand comprises a nucleotide sequence of any one of SEQ ID NOs: 819-849.77. The RNAi oligonucleotide of any one of clauses 1-59 and 75-76, wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of:(a) SEQ ID NOs: 774 and 819, respectively;(b) SEQ ID NOs: 775 and 820, respectively;(c) SEQ ID NOs: 776 and 821, respectively;(d) SEQ ID NOs: 777 and 822, respectively;(e) SEQ ID NOs: 778 and 823, respectively;(f) SEQ ID NOs: 779 and 824, respectively;(g) SEQ ID NOs: 780 and 825, respectively;(h) SEQ ID NOs: 781 and 826, respectively;(i) SEQ ID NOs: 782 and 827, respectively;(j) SEQ ID NOs: 783 and 828, respectively;(k) SEQ ID NOs: 784 and 829, respectively;(l) SEQ ID NOs: 785 and 830, respectively;(m) SEQ ID NOs: 786 and 831, respectively;(n) SEQ ID NOs: 787 and 832, respectively;(o) SEQ ID NOs: 788 and 833, respectively;(p) SEQ ID NOs: 789 and 834, respectively;(q) SEQ ID NOs: 790 and 835, respectively;(r) SEQ ID NOs: 791 and 836, respectively;(s) SEQ ID NOs: 792 and 837, respectively;(t) SEQ ID NOs: 793 and 838, respectively;(u) SEQ ID NOs: 794 and 839, respectively;(v) SEQ ID NOs: 795 and 840, respectively;(w) SEQ ID NOs: 796 and 841, respectively;(x) SEQ ID NOs: 797 and 842, respectively;(y) SEQ ID NOs: 798 and 843, respectively;(z) SEQ ID NOs: 799 and 844, respectively;(aa) SEQ ID NOs: 800 and 845, respectively;(bb) SEQ ID NOs: 801 and 846, respectively;(cc) SEQ ID NOs: 802 and 847, respectively;(dd) SEQ ID NOs: 803 and 848, respectively; and(ee) SEQ ID NOs: 804 and 849, respectively.78. The RNAi oligonucleotide of any one of clauses 1-59 and 75-76, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 804 and 849, respectively.79. The RNAi oligonucleotide of any one of clauses 1-59 and 75-76, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 782 and 827, respectively.80. The RNAi oligonucleotide of any one of clauses 1-59 and 75-76, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 775 and 820, respectively.81. The RNAi oligonucleotide of any one of clauses 1-59 and 75-76, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 779 and 824, respectively.82. The RNAi oligonucleotide of any one of clauses 1-59 and 75-76, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 780 and 825, respectively.83. The RNAi oligonucleotide of any one of clauses 1-59 and 75-76, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 785 and 830, respectively.84. The RNAi oligonucleotide of any one of clauses 1-59, wherein the sense strand comprises a nucleotide sequence of any one of SEQ ID NOs: 805-818.85. The RNAi oligonucleotide of any one of clauses 1-59 and 84, wherein the antisense strand comprises a nucleotide sequence of any one of SEQ ID NOs: 850-86386. The RNAi oligonucleotide of any one of clauses 1-59 and 84-85, wherein the sense and antisense strands comprise nucleotide sequences selected from the group consisting of:(a) SEQ ID NOs: 805 and 850, respectively;(b) SEQ ID NOs: 806 and 851, respectively;(c) SEQ ID NOs: 807 and 852, respectively;(d) SEQ ID NOs: 808 and 853, respectively;(e) SEQ ID NOs: 809 and 854, respectively;(f) SEQ ID NOs: 810 and 855, respectively;(g) SEQ ID NOs: 811 and 856, respectively;(h) SEQ ID NOs: 812 and 857, respectively;(i) SEQ ID NOs: 813 and 858, respectively;(j) SEQ ID NOs: 814 and 859, respectively;(k) SEQ ID NOs: 815 and 860, respectively;(l) SEQ ID NOs: 816 and 861, respectively;(m) SEQ ID NOs: 817 and 862, respectively and;(n) SEQ ID NOs: 818 and 863, respectively.87. The RNAi oligonucleotide of any one of clauses 1-59 and 84-86, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 805 and 850, respectively.88. The RNAi oligonucleotide of any one of clauses 1-59 and 84-86, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 809 and 854, respectively.89. The RNAi oligonucleotide of any one of clauses 1-59 and 84-86, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 810 and 855, respectively.90. The RNAi oligonucleotide of any one of clauses 1-59 and 84-86, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 812 and 857, respectively.91. The RNAi oligonucleotide of any one of clauses 1-59 and 84-86, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 815 and 860, respectively.92. The RNAi oligonucleotide of any one of clauses 1-59 and 84-86, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 818 and 863, respectively.93. A double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mG-S-mC-mA-mG-mG-mA-mA-fG-fC-fA-fC-mU-mG-mA-mG-mA-mU-mU-mC-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 804), and wherein the antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fG-S-fA-S-fA-fU-mC-fU-mC-mA-fG-mU-mG-mC-fU-mU-mC-mC-mU-mG-mC-S-mG-S-mG-3′ (SEQ ID NO: 849), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.94. A double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mU-S-mU-mU-mG-mA-mG-mA-fA-fG-fG-fU-mU-mG-mA-mU-mC-mU-mG-mA-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 782), and wherein the antisense strand comprises the sequence and all of the modifications of 5′ [MePhosphonate-4O-mU]-S-fU-S-fC-S-fA-fG-mA-fU-mC-mA-fA-mC-mC-mU-fU-mC-mU-mC-mA-mA-mA-S-mG-S-mG-3′ (SEQ ID NO: 827), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.95. A double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mG-S-mA-mA-mG-mA-mG-mA-fA-fG-fC-fA-mG-mA-mU-mC-mC-mU-mG-mU-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 775), and wherein the antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fA-S-fC-fA-fG-mG-fA-mU-mC-fU-mG-mC-mU-fU-mC-mU-mC-mU-mU-mC-S-mG-S-mG-3′ (SEQ ID NO: 820), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.96. A double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mC-S-mA-mG-mA-mU-mG-mU-mG-fU-fC-fU-mG-mC-mU-mA-mC-mA-mG-mA-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 779), and wherein the antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fU-S-fC-S-fU-fG-mU-fA-mG-mC-fA-mG-mA-mC-fA-mC-mA-mU-mC-mU-mG-S-mG-S-mG-3′ (SEQ ID NO: 824), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.97. A double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mG-S-mA-mC-mU-mU-mU-mG-fA-fG-fA-fA-mG-mG-mU-mU-mG-mA-mU-mC-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 780), and wherein the antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fG-S-fA-S-fU-fC-mA-fA-mC-mC-fU-mU-mC-mU-fC-mA-mA-mA-mG-mU-mC-S-mG-S-mG-3′ (SEQ ID NO: 825), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.98. A double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein the sense strand comprises the sequence and all of the modifications of 5′-mU-S-mG-mU-mU-mU-mG-mU-fC-fA-fG-fC-mA-mA-mA-mG-mA-mU-mG-mU-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 785), and wherein the antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fA-S-fC-fA-fU-mC-fU-mU-mU-fG-mC-mU-mG-fA-mC-mA-mA-mA-mC-mA-S-mG-S-mG-3′ (SEQ ID NO: 830), wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

or a pharmaceutically acceptable salt thereof.99. A dsRNAi oligonucleotide for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand comprising SEQ ID NO: 775 and an antisense strand comprising SEQ ID NO: 820, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure as depicted in FIG. 10A continuing to FIG. 10B, or a pharmaceutically acceptable salt thereof.100. A dsRNAi oligonucleotide for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand comprising SEQ ID NO: 779 and an antisense strand comprising SEQ ID NO: 824, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure as depicted FIG. 11A continuing to FIG. 11B, or a pharmaceutically acceptable salt thereof.101. A dsRNAi oligonucleotide for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand comprising SEQ ID NO: 780 and an antisense strand comprising SEQ ID NO: 825, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure as depicted in FIG. 12A continuing to FIG. 12B, or a pharmaceutically acceptable salt thereof.102. A dsRNAi oligonucleotide for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand comprising SEQ ID NO: 782 and an antisense strand comprising SEQ ID NO: 827, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure FIG. 13A continuing to FIG. 13B, or a pharmaceutically acceptable salt thereof.103. A dsRNAi oligonucleotide for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand comprising SEQ ID NO: 785 and an antisense strand comprising SEQ ID NO: 830, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure FIG. 14A continuing to FIG. 14B, or a pharmaceutically acceptable salt thereof.104. A dsRNAi oligonucleotide for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand comprising SEQ ID NO: 804 and an antisense strand comprising SEQ ID NO: 849, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure of FIG. 15A continuing to FIG. 15B, or a pharmaceutically acceptable salt thereof.105. The RNAi oligonucleotide of any one of clauses 1-104, wherein expression of KHK is reduced or inhibited in vivo.106. The RNAi oligonucleotide of any one of clauses 1-105, wherein the oligonucleotide is a Dicer substrate.107. The RNAi oligonucleotide of any one of clauses 1-105, wherein the oligonucleotide is a Dicer substrate that, upon endogenous Dicer processing, yields double-stranded nucleic acids of 19-23 nucleotides in length capable of reducing KHK expression in a mammalian cell.108. A cell containing the RNAi oligonucleotide of any one of the preceding clauses.109. A method for treating a subject having a disease, disorder or condition associated with KHK expression, the method comprising administering to the subject a therapeutically effective amount of the RNAi oligonucleotide of any one of clauses 1-107, or pharmaceutically acceptable salt thereof, or pharmaceutical composition thereof, thereby treating the subject.110. A pharmaceutical composition comprising the RNAi oligonucleotide of any one of clauses 1-107, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, delivery agent or excipient.111. A method of delivering an oligonucleotide to a subject, the method comprising administering the pharmaceutical composition of clause 110 to the subject.112. An in vitro or in vivo method for modulating, e.g. inhibiting or reducing, KHK expression in a target cell expressing KHK, the method comprising administering the pharmaceutical composition of clause 110 in an effective amount to the target cell.113. A method for reducing KHK expression in a cell, a population of cells or a subject, the method comprising the step of:

i. contacting the cell or the population of cells with the RNAi oligonucleotide, or a pharmaceutically acceptable salt thereof, of any one of clauses 1-107, or the pharmaceutical composition of clause 110; or

ii. administering to the subject the RNAi oligonucleotide, or a pharmaceutically acceptable salt thereof of any one of clauses 1-107, or the pharmaceutical composition of clause 110.

114. The method of clause 113, wherein reducing KHK expression comprises reducing an amount or level of KHK mRNA, an amount or level of KHK protein, or both.115. The method of any one of clauses 111 and 113-114, wherein the subject has a disease, disorder or condition associated with KHK expression.116. The method of clause 115, wherein the disease, disorder or condition associated with KHK expression is non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).117. The method of any one of clauses 109 and 111-116, wherein the RNAi oligonucleotide, or pharmaceutical composition, is administered in combination with a second composition or therapeutic agent.118. A method for treating a subject having a disease, disorder or condition associated with KHK expression, the method comprising administering to the subject a therapeutically effective amount of an RNAi oligonucleotide comprising a sense strand and an antisense strand, or a pharmaceutically acceptable salt thereof, wherein the sense strand and antisense strand comprise nucleotide sequences selected from the group consisting of:

• • (a) SEQ ID NOs: 886 and 912, respectively;
  • (b) SEQ ID NOs: 887 and 913, respectively;
  • (c) SEQ ID NOs: 910 and 937, respectively;
  • (d) SEQ ID NOs: 888 and 914, respectively;
  • (e) SEQ ID NOs: 889 and 915, respectively;
  • (f) SEQ ID NOs: 890 and 916, respectively;
  • (g) SEQ ID NOs: 891 and 917, respectively;
  • (h) SEQ ID NOs: 877 and 884, respectively;
  • (i) SEQ ID NOs: 878 and 930, respectively;
  • (j) SEQ ID NOs: 876 and 883, respectively;
  • (k) SEQ ID NOs: 875 and 882, respectively;
  • (l) SEQ ID NOs: 892 and 918, respectively;
  • (m) SEQ ID NOs: 893 and 919, respectively;
  • (n) SEQ ID NOs: 894 and 920, respectively;
  • (o) SEQ ID NOs: 904 and 931, respectively;
  • (p) SEQ ID NOs: 895 and 921, respectively;
  • (q) SEQ ID NOs: 905 and 932, respectively;
  • (r) SEQ ID NOs: 896 and 922, respectively;
  • (s) SEQ ID NOs: 911 and 938, respectively;
  • (t) SEQ ID NOs: 906 and 933, respectively;
  • (u) SEQ ID NOs: 897 and 923, respectively;
  • (v) SEQ ID NOs: 907 and 934, respectively;
  • (w) SEQ ID NOs: 908 and 935, respectively;
  • (x) SEQ ID NOs: 903 and 929, respectively;
  • (y) SEQ ID NOs: 901 and 927, respectively;
  • (z) SEQ ID NOs: 874 and 881, respectively;
  • (aa) SEQ ID NOs: 902 and 928, respectively;
  • (bb) SEQ ID NOs: 873 and 880, respectively;
  • (cc) SEQ ID NOs: 872 and 879, respectively;
  • (dd) SEQ ID NOs: 898 and 924, respectively;
  • (ee) SEQ ID NOs: 899 and 925, respectively (ff) SEQ ID NOs: 900 and 926, respectively; and
  • (gg) SEQ ID NOs: 909 and 936, respectively.119. The method of clause 118, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 909 and 936, respectively.120. The method of clause 118, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 894 and 920, respectively.121. The method of clause 118, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 897 and 923, respectively.122. The method of clause 118, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 892 and 918, respectively.123. The method of clause 118, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 891 and 917, respectively.124. The method of clause 118, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 887 and 913, respectively.125. A method for treating a subject having a disease, disorder or condition associated with KHK expression, the method comprising administering to the subject a therapeutically effective amount of an RNAi oligonucleotide comprising a sense strand and an antisense strand, or a pharmaceutically acceptable salt thereof, wherein the sense strand and antisense strands are selected from the group consisting of:(a) SEQ ID NOs: 774 and 819, respectively;(b) SEQ ID NOs: 775 and 820, respectively;(c) SEQ ID NOs: 776 and 821, respectively;(d) SEQ ID NOs: 777 and 822, respectively;(e) SEQ ID NOs: 778 and 823, respectively;(f) SEQ ID NOs: 779 and 824, respectively;(g) SEQ ID NOs: 780 and 825, respectively;(h) SEQ ID NOs: 781 and 826, respectively;(i) SEQ ID NOs: 782 and 827, respectively;(j) SEQ ID NOs: 783 and 828, respectively;(k) SEQ ID NOs: 784 and 829, respectively;(l) SEQ ID NOs: 785 and 830, respectively;(m) SEQ ID NOs: 786 and 831, respectively;(n) SEQ ID NOs: 787 and 832, respectively;(o) SEQ ID NOs: 788 and 833, respectively;(p) SEQ ID NOs: 789 and 834, respectively;(q) SEQ ID NOs: 790 and 835, respectively;(r) SEQ ID NOs: 791 and 836, respectively;(s) SEQ ID NOs: 792 and 837, respectively;(t) SEQ ID NOs: 793 and 838, respectively;(u) SEQ ID NOs: 794 and 839, respectively;(v) SEQ ID NOs: 795 and 840, respectively;(w) SEQ ID NOs: 796 and 841, respectively;(x) SEQ ID NOs: 797 and 842, respectively;(y) SEQ ID NOs: 798 and 843, respectively;(z) SEQ ID NOs: 799 and 844, respectively;(aa) SEQ ID NOs: 800 and 845, respectively;(bb) SEQ ID NOs: 801 and 846, respectively;(cc) SEQ ID NOs: 802 and 847, respectively;(dd) SEQ ID NOs: 803 and 848, respectively; and(ee) SEQ ID NOs: 804 and 849, respectively.126. The method of clause 125, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 804 and 849, respectively.127. The method of clause 125 wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 782 and 827, respectively.128. The method of clause 125, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 775 and 820, respectively.129. The method of clause 125, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 779 and 824, respectively.130. The method of clause 125, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 780 and 825, respectively.131. The method of clause 125, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 785 and 830, respectively.132. A method for treating a subject having a disease, disorder or condition associated with KHK expression, the method comprising administering to the subject a therapeutically effective amount of an RNAi oligonucleotide comprising a sense strand and an antisense strand, or a pharmaceutically acceptable salt thereof, wherein the sense strand and antisense strands are selected from the group consisting of:(a) SEQ ID NOs: 805 and 850, respectively;(b) SEQ ID NOs: 806 and 851, respectively;(c) SEQ ID NOs: 807 and 852, respectively;(d) SEQ ID NOs: 808 and 853, respectively;(e) SEQ ID NOs: 809 and 854, respectively;(f) SEQ ID NOs: 810 and 855, respectively;(g) SEQ ID NOs: 811 and 856, respectively;(h) SEQ ID NOs: 812 and 857, respectively;(i) SEQ ID NOs: 813 and 858, respectively;(j) SEQ ID NOs: 814 and 859, respectively;(k) SEQ ID NOs: 815 and 860, respectively;(l) SEQ ID NOs: 816 and 861, respectively;(m) SEQ ID NOs: 817 and 862, respectively and;(n) SEQ ID NOs: 818 and 863, respectively.133. The method of clause 132, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 805 and 850, respectively.134. The method of clause 132, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 809 and 854, respectively.135. The method of clause 132, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 810 and 855, respectively.136. The method of clause 132, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 812 and 857, respectively.137. The method of clause 132, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 815 and 860, respectively.138. The method of clause 132, wherein the sense and antisense strands comprise the nucleotide sequences set forth in SEQ ID NOs: 818 and 863, respectively.139. The method of any one of clauses 118-138, wherein the disease, disorder or condition associated with KHK expression is non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).140. The method of any one of clauses 109, 111 and 113-132, wherein the dsRNA is administered at a concentration of 0.01 mg/kg-5 mg/kg bodyweight of the subject.141. Use of the RNAi oligonucleotide of any one of clauses 1-107, or the pharmaceutical composition of clause 110, in the manufacture of a medicament for the treatment of a disease, disorder or condition associated with KHK expression, optionally for the treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).142. The RNAi oligonucleotide of any one of clauses 1-107, or the pharmaceutical composition of clause 110, for use, or adaptable for use, in the treatment of a disease, disorder or condition associated with KHK expression, optionally for the treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).143. A kit comprising the RNAi oligonucleotide of any one of clauses 1-107, an optional pharmaceutically acceptable carrier, and a package insert comprising instructions for administration to a subject having a disease, disorder or condition associated with KHK expression.144. The use of clause 141, the RNAi oligonucleotide or pharmaceutical composition for use, or adaptable for use, of clause 142, or the kit of clause 143, wherein the disease, disorder or condition associated with KHK expression is non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).145. An oligonucleotide for reducing KHK expression, the oligonucleotide comprising a nucleotide sequence of 15-50 nucleotides in length, wherein the nucleotide sequence comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387, and wherein the region of complementarity is at least 15 contiguous nucleotides.146. The oligonucleotide of clause 145, wherein the oligonucleotide is single stranded.147. The oligonucleotide of clause 145 or 146, wherein the oligonucleotide is an antisense oligonucleotide.148. The oligonucleotide of any one of clauses 145-147, wherein the nucleotide sequence is 15-30 nucleotides in length.149. The oligonucleotide of any one of clauses 145-148, wherein the nucleotide sequence is 20-25 nucleotides in length.150. The oligonucleotide of any one of clauses 145-149, wherein the nucleotide sequence is 22 nucleotides in length.151. The oligonucleotide of any one of clauses 145-150, wherein the region of complementarity is 19 contiguous nucleotides in length.152. The oligonucleotide of any one of clauses 145-150, wherein the region of complementarity is 20 contiguous nucleotides in length.153. The oligonucleotide of any one of clauses 145-152, wherein the nucleotide sequence comprises at least one modification.154. The oligonucleotide of any one of clauses 145-153, wherein the nucleotide sequence comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 879-884 and 912-938.155. The oligonucleotide of any one of clauses 145-153, wherein the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 909.156. The oligonucleotide of any one of clauses 145-153, wherein the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 894.157. The oligonucleotide of any one of clauses 145-153, wherein the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 897.158. The oligonucleotide of any one of clauses 145-153, wherein the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 892.159. The oligonucleotide of any one of clauses 145-153, wherein the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 891.160. The oligonucleotide of any one of clauses 145-153, wherein the nucleotide sequence comprises the nucleotide sequence set forth in SEQ ID NO: 887.161. A cell comprising the oligonucleotide of any one of clauses 145-160. 162. A pharmaceutical composition comprising the oligonucleotide of any one of clauses 145-160, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, delivery agent or excipient.163. A method for treating a subject having a disease, disorder or condition associated with

KHK expression, the method comprising administering to the subject a therapeutically effective amount of the oligonucleotide of any one of clauses 145-160, or pharmaceutical composition of clause 162.

164. A method of delivering an oligonucleotide to a subject, the method comprising administering the pharmaceutical composition of clause 162 to the subject.165. A method for reducing KHK expression in a cell, a population of cells or a subject, the method comprising the step of:

i. contacting the cell or the population of cells with the oligonucleotide of any one of clauses 145-160, or the pharmaceutical composition of clause 162; or

ii. administering to the subject the oligonucleotide of any one of clauses 145-160, or the pharmaceutical composition of clause 162.

166. The method of clause 165, wherein reducing KHK expression comprises reducing an amount or level of KHK mRNA, an amount or level of KHK protein, or both.167. The method of any one of clauses 164-166, wherein the subject has a disease, disorder or condition associated with KHK expression.168. The method of clause 167, wherein the disease, disorder or condition associated with KHK expression is non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).169. The method of any one of clauses 163-168, wherein the oligonucleotide, or pharmaceutical composition, is administered in combination with a second composition or therapeutic agent.170. Use of the oligonucleotide of any one of clauses 145-160, or the pharmaceutical composition of clause 161, in the manufacture of a medicament for the treatment of a disease, disorder or condition associated with KHK expression, optionally for the treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).171. The oligonucleotide of any one of clauses 145-160, or the pharmaceutical composition of clause 161, for use, or adaptable for use, in the treatment of a disease, disorder or condition associated with KHK expression, optionally for the treatment of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).172. A kit comprising the oligonucleotide of any one of clauses 145-160, an optional pharmaceutically acceptable carrier, and a package insert comprising instructions for administration to a subject having a disease, disorder or condition associated with KHK expression.173. The use of clause 170, the RNAi oligonucleotide or pharmaceutical composition for use, or adaptable for use, of clause 171, or the kit of clause 172, wherein the disease, disorder or condition associated with KHK expression is non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).174. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of KHK, wherein the dsRNA agent comprises a sense strand and an antisense strand forming a duplex region, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from a nucleotide sequence selected from SEQ ID NOs: 4-387, and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from a nucleotide sequence selected from SEQ ID NOs: 388-771, or a pharmaceutically acceptable salt thereof.175. A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of KHK, wherein the dsRNA agent comprises a sense strand and an antisense strand forming a duplex region, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from a nucleotide sequence selected from SEQ ID NOs: 872-878 and 886-911, and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from a nucleotide sequence selected from SEQ ID NOs: 879-884 and 912-938, or a pharmaceutically acceptable salt thereof.176. A pharmaceutical composition comprising the dsRNA agent of clause 174 or 175, and a pharmaceutically acceptable diluent, solvent, carrier, salt, and/or adjuvant.177. An in vitro or in vivo method for reducing or inhibiting KHK expression in a target cell expressing KHK, the method comprising administering the pharmaceutical composition of clause 176 in an effective amount to the target cell.178. A method for treating or preventing a disease associated with KHK expression, comprising administering a therapeutically or prophylactically effective amount of the pharmaceutical composition of clause 176 to a subject suffering from or susceptible to the disease.179. The method of any one of clauses 109 and 113-140, wherein a single dose of one or more RNAi oligonucleotides of any one of clauses 1-107, or pharmaceutically acceptable salts thereof, or the pharmaceutical composition of any one of clauses 110, 162, or 176 is administered such that an amount or level of KHK mRNA and/or KHK protein is reduced in the subject when compared to KHK expression prior to administration of the one or more RNAi oligonucleotides, or pharmaceutically acceptable salts thereof, or the pharmaceutical composition and/or when compared to KHK expression in a subject not receiving the one or more RNAi oligonucleotides, or pharmaceutically acceptable salts thereof, or pharmaceutical composition or receiving one or more control oligonucleotides, pharmaceutical compositions or treatments, and wherein said reduction remains detectable at day 28, 56, and/or 84 after the single dose administration.180. The method of clause 179, wherein the amount or level of KHK mRNA and/or KHK protein is reduced by at least about 30%, by at least about 50%, or by at least about 70%. 181. The method of any one of clauses 179-180, wherein the dose is administered subcutaneously.

Claims

1. A double stranded RNAi oligonucleotide for reducing ketohexokinase (KHK) expression, the oligonucleotide comprising an antisense strand and a sense strand, wherein the antisense strand and the sense strand form a duplex region, wherein the antisense strand comprises a region of complementarity to a KHK mRNA target sequence of any one of SEQ ID NOs: 4-387 and wherein the region of complementarity is at least 15 contiguous nucleotides in length, or a pharmaceutically acceptable salt thereof, wherein the sense strand comprises a sequence set forth in any one of SEQ ID NOs: 4-387 and/orthe antisense strand comprises a sequence set forth in any one of SEQ ID NOs: 388-771.

2. A double stranded RNAi oligonucleotide for inhibiting expression of KHK, wherein said double stranded RNAi oligonucleotide comprises a sense strand and an antisense strand forming a duplex region, wherein said sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the nucleotide sequences of SEQ ID NO:4-387 and said antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the nucleotide sequences of SEQ ID NO: 388-771, or a pharmaceutically acceptable salt thereof, wherein the sense strand is 18 to 36 nucleotides in length and/or the antisense strand is 15-30 nucleotides in length.

3. A double stranded RNAi (dsRNAi) oligonucleotide for reducing or inhibiting ketohexokinase (KHK) expression, the oligonucleotide comprising: (i) an antisense strand of 19-30 nucleotides in length, wherein the antisense strand comprises a nucleotide sequence comprising a region of complementarity to a KHK mRNA target sequence, wherein the region of complementarity is selected from SEQ ID NOs: 948-953; and(ii) a sense strand of 19-50 nucleotides in length comprising a region of complementarity to the antisense strand,wherein the antisense and sense strands are separate strands which form an asymmetric duplex region having an overhang of 1-4 nucleotides at the 3′ terminus of the antisense strand.

4. The RNAi oligonucleotide of claim 1, wherein the region of complementarity comprised by the antisense strand is at least 19 contiguous nucleotides in length.

5. The RNAi oligonucleotide of claim 1, wherein the duplex region is at least 20 nucleotides in length.

6. The RNAi oligonucleotide of claim 1, wherein the sense strand comprises at its 3′ end a stem-loop set forth as: S1-L-S2, wherein S1 is complementary to S2, andwherein L forms a loop between S1 and S2 of 3 to 5 nucleotides in length,preferably the stem-loop comprises the sequence 5′-GCAGCCGAAAGGCUGC-3′ (SEQ ID NO: 871).

7. The RNAi oligonucleotide of claim 6, wherein at least one nucleotide of the oligonucleotide is conjugated to one or more targeting ligands,wherein each targeting ligand comprises an N-acetylgalactosamine (GalNAc) moiety,wherein the one or more targeting ligands is conjugated to one or more nucleotides of the loop according to claim 6.

8. The RNAi oligonucleotide of claim 1, wherein the overhang is 2 nucleotides in length and is selected from AA, GG, AG, and GA.

9. The RNAi oligonucleotide of any one of claim 1, wherein all the nucleotides of the oligonucleotide are modified, 10-15%, 10%, 11%, 12%, 13%, 14% or 15% of the nucleotides of the sense strand comprise a 2′-fluoro modificationand/or about 25-35%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34% or 35% of the nucleotides of the antisense strand comprise a 2′-fluoro modification.

10. The RNAi oligonucleotide of claim 1, wherein the oligonucleotide comprises at least one phosphorothioate linkage between positions 1 and 2, between positions 2 and 3, and between positions 3 and 4,wherein positions are numbered 1-4 from 5′ to 3′.

11. The RNAi oligonucleotide of claim 1, wherein the 4′-carbon of the sugar of the 5′-terminal nucleotide of the antisense strand comprises a 4′-phosphate analog comprising 5′-methoxyphosphonate-4′-oxy.

12. The RNAi oligonucleotide of claim 1, wherein the antisense strand is 22 nucleotides in length and/orwherein the sense strand is 36 nucleotides in length.

13. The RNAi oligonucleotide of claim 1, wherein the sense and antisense strands comprise the nucleotide sequences set forth in(a) SEQ ID NOs: 887 and 913, respectively, or(b) SEQ ID NOs: 891 and 917, respectively, or(c) SEQ ID NOs: 892 and 918, respectively, or(d) SEQ ID NOs: 894 and 920, respectively, or(e) SEQ ID NOs: 897 and 923, respectively, or(f) SEQ ID NOs: 909 and 936, respectively.

14. A double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNAi comprises a sense strand and an antisense strand, the antisense strand comprising a region of complementarity to a KHK RNA transcript, wherein (a) the sense strand comprises the sequence and all of the modifications of 5′-mG-S-mA-mA-mG-mA-mG-mA-fA-fG-fC-fA-mG-mA-mU-mC-mC-mU-mG-mU-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 775), andthe antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fA-S-fC-fA-fG-mG-fA-mU-mC-fU-mG-mC-mU-fU-mC-mU-mC-mU-mU-mC-S-mG-S-mG-3′ (SEQ ID NO: 820); or(b) the sense strand comprises the sequence and all of the modifications of 5′-mC-S-mA-mG-mA-mU-mG-mU-fG-fU-fC-fU-mG-mC-mU-mA-mC-mA-mG-mA-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 779), andthe antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fU-S-fC-S-fU-fG-mU-fA-mG-mC-fA-mG-mA-mC-fA-mC-mA-mU-mC-mU-mG-S-mG-S-mG-3′ (SEQ ID NO: 824); or(c) the sense strand comprises the sequence and all of the modifications of 5′-mG-S-mA-mC-mU-mU-mU-mG-fA-fG-fA-fA-mG-mG-mU-mU-mG-mA-mU-mC-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 780), andthe antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fG-S-fA-S-fU-fC-mA-fA-mC-mC-fU-mU-mC-mU-fC-mA-mA-mA-mG-mU-mC-S-mG-S-mG-3′ (SEQ ID NO: 825); or(d) the sense strand comprises the sequence and all of the modifications of 5′-mU-S-mU-mU-mG-mA-mG-mA-fA-fG-fG-fU-mU-mG-mA-mU-mC-mU-mG-mA-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 782), andthe antisense strand comprises the sequence and all of the modifications of 5′ [MePhosphonate-4O-mU]-S-fU-S-fC-S-fA-fG-mA-fU-mC-mA-fA-mC-mC-mU-fU-mC-mU-mC-mA-mA-mA-S-mG-S-mG-3′ (SEQ ID NO: 827); or(e) the sense strand comprises the sequence and all of the modifications of 5′-mU-S-mG-mU-mU-mU-mG-mU-fC-fA-fG-fC-mA-mA-mA-mG-mA-mU-mG-mU-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 785), andthe antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fA-S-fC-fA-fU-mC-fU-mU-mU-fG-mC-mU-mG-fA-mC-mA-mA-mA-mC-mA-S-mG-S-mG-3′ (SEQ ID NO: 830); or(f) the sense strand comprises the sequence and all of the modifications of 5′-mG-S-mC-mA-mG-mG-mA-mA-fG-fC-fA-fC-mU-mG-mA-mG-mA-mU-mU-mC-mA-mG-mC-mA-mG-mC-mC-mG-[ademA-GalNAc]-[ademA-GalNAc]-[ademA-GalNAc]-mG-mG-mC-mU-mG-mC-3′ (SEQ ID NO: 804), andthe antisense strand comprises the sequence and all of the modifications of 5′-[MePhosphonate-4O-mU]-S-fG-S-fA-S-fA-fU-mC-fU-mC-mA-fG-mU-mG-mC-fU-mU-mC-mC-mU-mG-mC-S-mG-S-mG-3′ (SEQ ID NO: 849);wherein mC, mA, mG, mU=2′-OMe ribonucleosides; fA, fC, fG, fU=2′F ribonucleosides; “-”=phosphodiester linkage, “—S—”=phosphorothioate linkage, and wherein ademA-GalNAc=

15. A double stranded RNAi oligonucleotide (dsRNAi) for inhibiting expression of KHK, wherein said dsRNAi comprises (a) a sense strand comprising SEQ ID NO: 775 and an antisense strand comprising SEQ ID NO: 820, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure depicted in FIG. 10A continuing to FIG. 10B;or(b) a sense strand comprising SEQ ID NO: 779 and an antisense strand comprising SEQ ID NO: 824, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure depicted in FIG. 11A continuing to FIG. 11B;or(c) a sense strand comprising SEQ ID NO: 780 and an antisense strand comprising SEQ ID NO: 825, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure depicted in depicted in FIG. 12A continuing to FIG. 12B;or(d) a sense strand comprising SEQ ID NO: 782 and an antisense strand comprising SEQ ID NO: 827, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure depicted in FIG. 13A continuing to FIG. 13B;or(e) a sense strand comprising SEQ ID NO: 785 and an antisense strand comprising SEQ ID NO: 830, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure depicted in FIG. 14A continuing to FIG. 14B;or(f) a sense strand comprising SEQ ID NO: 804 and an antisense strand comprising SEQ ID NO: 849, the antisense strand comprising a region of complementarity to a KHK RNA transcript, e.g. KHK mRNA, wherein said dsRNAi is in the form of a conjugate having the structure depicted in FIG. 15A continuing to FIG. 15B;or a pharmaceutically acceptable salt thereof.

16. A pharmaceutical composition comprising the dsRNAi oligonucleotide of claim 1, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, delivery agent or excipient.

17. A method for treating a disease, disorder or condition associated with KHK expression, comprising administering to a patient in need thereof a pharmaceutically effective amount of the RNAi oligonucleotide of claim 1.

18. The method of claim 14, wherein the disease, disorder or condition is selected from the group consisting of non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH).

19. The method according to claim 18, further comprising administering the RNAi in combination with a second therapeutic agent.

20. A method for reducing KHK expression in a cell, a population of cells or a subject, the method comprising the step of: i. contacting the cell or the population of cells with the RNAi oligonucleotide, or a pharmaceutically acceptable salt thereof, of claim 1; orii. administering to the subject the RNAi oligonucleotide, or a pharmaceutically acceptable salt thereof of claim 1.