Methods of Modulating the Sex of Avians

Abstract
The present invention relates to nucleic acids and methods for modulating the sex of avians. In particular, the invention relates to the in ovo delivery of a dsRNA molecule, especially siRNAs, to increase the production of female birds.
Description
FIELD OF THE INVENTION

The present invention relates to nucleic acids and methods for modulating the sex of avians. In particular, the invention relates to the in ovo delivery of a dsRNA molecule, especially siRNAs, to increase the production of female birds.


BACKGROUND OF THE INVENTION

Man has modified the phenotypic characteristics of domestic animals through selection of seed stock over many generations ever since animals were domesticated. This has led to improvements in quantitative production parameters such as body size and muscle mass. More recent innovations of modifying production traits of poultry and/or improving resistance to pathogens has focussed on transgenic approaches, however, many consumers have concerns about genetically modified organisms.


Chicken producers have been searching for an efficient, economical method of determining the sex of day old chicks. Vent sexing and feather sexing have been used by the various producers, but these methods have been found to have substantial economic disadvantages because of the substantial time required and labour costs in separating the male from the female chicks. The use of probes (U.S. Pat. No. 5,508,165) is also an expensive procedure and not practical economically. Light sensing of anal areas of chicks (U.S. Pat. No. 4,417,663) is another way of determining sex of chicks, but it is also expensive and time consuming as each chick must be handled and manipulated. The use of experts who could feather sex the chicks has been used, but such experts are costly and feathering is time consuming.


There is a need for nucleic acids and methods for modifying avian sex that do not result in transformation of the bird's genome, but are amenable to high throughout processing.


SUMMARY OF THE INVENTION

The present inventors have identified nucleic acid molecules, in particular dsRNA molecules, which can be used to modify the sex of avians in ovo.


Accordingly, in one aspect the present invention provides an isolated and/or exogenous nucleic acid molecule comprising a double-stranded region which reduces the level of at least one RNA molecule and/or protein when administered to an avian egg, wherein if the embryo of the egg is male the sex is altered to female following administration of the isolated and/or exogenous nucleic acid molecule, and wherein the isolated and/or exogenous nucleic acid molecule does not comprise a sequence selected from:


CCAGUUGUCAAGAAGAGCA (SEQ ID NO:254)
GGAUGCUCAUUCAGGACAU (SEQ ID NO:369)
CCCUGUAUCCUUACUAUAA (SEQ ID NO:474)
GCCACUGAGUCUUCCUCAA (SEQ ID NO:530)
CCAGCAACAUACAUGUCAA (SEQ ID NO:605)
CCUGCGUCACACAGAUACU (SEQ ID NO:747)
GGAGUAGUUGUACAGGUUG (SEQ ID NO:3432)
GACUGGCUUGACAUGUAUG (SEQ ID NO:3433)

AUGGCGGUUCUCCAUCCCU (SEQ ID NO:3434) or a variant of any one thereof.


In another aspect, the present invention provides an isolated and/or exogenous nucleic acid molecule comprising one or more of the sequence of nucleotides provided as SEQ ID NO's 11 to 3431 or a variant of any one or more thereof, wherein the isolated and/or exogenous nucleic acid molecule does not comprise a sequence selected from:


CCAGUUGUCAAGAAGAGCA (SEQ ID NO:254)
GGAUGCUCAUUCAGGACAU (SEQ ID NO:369)
CCCUGUAUCCUUACUAUAA (SEQ ID NO:474)
GCCACUGAGUCUUCCUCAA (SEQ ID NO:530)
CCAGCAACAUACAUGUCAA (SEQ ID NO:605)
CCUGCGUCACACAGAUACU (SEQ ID NO:747)
GGAGUAGUUGUACAGGUUG (SEQ ID NO:3432)
GACUGGCUUGACAUGUAUG (SEQ ID NO:3433)

AUGGCGGUUCUCCAUCCCU (SEQ ID NO:3434) or a variant of any one thereof.


In a preferred embodiment, the nucleic acid molecule is dsRNA. More preferably, the dsRNA is a siRNA or a shRNA.


In a preferred embodiment, the nucleic acid molecule reduces the level of a protein encoded by a DMRT1 gene, ASW gene or r-spondin gene, in an avian egg.


In a preferred embodiment of the two above aspects, the nucleic acid does not comprise the full length open reading frame of the RNA molecule or the cDNA encoding therefor. In a related embodiment, preferably the nucleic acid does not comprise a sequence of nucleotides provided as SEQ ID NO's 2, 4 or 6, or a sequence of nucleotides provided as SEQ ID NO's 2, 4 or 6 where each T (thymine) is replaced with a U (uracil).


As the skilled addressee will appreciate, because the nucleic acid is double stranded it will also comprise the corresponding reverse complement of the relevant nucleotide sequence provided herewith.


Also provided is a vector encoding a nucleic acid molecule, or a single strand thereof, according to the invention. Such vectors can be used in a host cell or cell-free expression system to produce nucleic acid molecules useful for the method of the invention.


In another aspect, the present invention provides a host cell comprising an exogenous nucleic acid molecule, or a single strand thereof, of the invention and/or a vector of the invention.


In another aspect, the present invention provides a composition comprising a nucleic acid molecule, or a single strand thereof, of the invention, a vector of the invention, and/or a host cell of the invention.


In a further aspect, the present invention provides a method of modifying the sex of an avian, the method comprising administering to an avian egg at least one nucleic acid molecule of the invention.


Preferably, the nucleic acid is administered to a non-cellular site of the egg. More preferably, the non-cellular site is the air sac, yolk sac, amnionic cavity or chorion allantoic fluid.


In a further preferred embodiment, the egg is not electroporated.


Preferably, the nucleic acid is not delivered by administering a vector encoding the nucleic acid molecule.


Preferably, the nucleic acid molecule administered is dsRNA.


Preferably, the nucleic acid molecule is administered by injection.


Conveniently, the nucleic acid may be administered in a composition of the invention.


In a preferred embodiment, the method modifies the sex of the embryo of the egg from male to female.


The avian can be any species of the Class Ayes. Examples include, but are not limited to, chickens, ducks, turkeys, geese, bantams and quails. In a particularly preferred embodiment, the avian is a chicken.


In a further aspect, the present invention provides an avian produced using a method of the invention.


In another aspect, the present invention provides a chicken produced using a method of the invention.


In a further aspect, the present invention provides an avian egg comprising a nucleic acid molecule, or a single strand thereof, of the invention, a vector of the invention, and/or a host cell of the invention.


In another aspect, the present invention provides a kit comprising a nucleic acid molecule, or a single strand thereof, of the invention, a vector of the invention, a host cell of the invention, and/or a composition of the invention.


As will be apparent, preferred features and characteristics of one aspect of the invention are applicable to many other aspects of the invention.


Throughout this specification the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.


The invention is hereinafter described by way of the following non-limiting Examples and with reference to the accompanying figures.





BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS

FIG. 1—Selected shRNAs for knockdown of EGFP-Dmrt1 gene fusion expression. Mean fluorescence intensity for each transfection condition expressed relative to pEGFP-Dmrt1. Error bars indicate standard error calculated on each individual experiment completed in triplicate.


FIG. 2—qPCR analysis of DMRT1 gene expression following silencing.





KEY TO THE SEQUENCE LISTING

SEQ ID NO:1—Partial chicken DMRT1 protein sequence (Genbank AF123456).


SEQ ID NO:2—Partial nucleotide sequence encoding chicken DMRT1 (Genbank AF123456).


SEQ ID NO:3—Chicken WPKCI (ASW) (Genbank AF148455).

SEQ ID NO:4—Nucleotide sequence encoding chicken WPKCI (ASW) (Genbank AF148455).


SEQ ID NO:5—Chicken r-spondin (Genbank XM417760).


SEQ ID NO:6—Nucleotide sequence encoding chicken r-spondin (Genbank XM417760).


SEQ ID NO:7—Nucleotide sequence of chicken U6-1 promoter.


SEQ ID NO:8—Nucleotide sequence of chicken U6-3 promoter.


SEQ ID NO:9—Nucleotide sequence of chicken U6-4 promoter.


SEQ ID NO:10—Nucleotide sequence of chicken 7SK promoter.


SEQ ID NO's 11 to 3430—RNA sequences provided in Tables 1 to 3 for silencing the chicken DMRT1, ASW or r-spondin genes.


SEQ ID NO's 3431 to 3434—RNA sequences suitable for silencing the chicken DMRT1 gene.


SEQ ID NO's 3435 to 3485—Target regions of chicken DMRT1.


SEQ ID NO's 3486 to 3499—Oligonucleotide primers and probes.


DETAILED DESCRIPTION OF THE INVENTION
General Techniques and Definitions

Unless specifically defined otherwise, all technical and scientific terms used herein shall be taken to have the same meaning as commonly understood by one of ordinary skill in the art (e.g., in cell culture, molecular genetics, avian biology, RNA interference, and biochemistry).


Unless otherwise indicated, the recombinant protein, cell culture, and immunological techniques utilized in the present invention are standard procedures, well known to those skilled in the art. Such techniques are described and explained throughout the literature in sources such as, J. Perbal, A Practical Guide to Molecular Cloning, John Wiley and Sons (1984), J. Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbour Laboratory Press (1989), T. A. Brown (editor), Essential Molecular Biology: A Practical Approach, Volumes 1 and 2, IRL Press (1991), D. M. Glover and B. D. Hames (editors), DNA Cloning: A Practical Approach, Volumes 1-4, IRL Press (1995 and 1996), and F. M. Ausubel et al. (editors), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience (1988, including all updates until present), Ed Harlow and David Lane (editors).


The term “avian” as used herein refers to any species, subspecies or race of organism of the taxonomic Class Ayes, such as, but not limited to, such organisms as chicken, turkey, duck, goose, quail, pheasants, parrots, finches, hawks, crows and ratites including ostrich, emu and cassowary. The term includes the various known strains of Gallus gallus (chickens), for example, White Leghorn, Brown Leghorn, Barred-Rock, Sussex, New Hampshire, Rhode Island, Australorp, Cornish, Minorca, Amrox, California Gray, Italian Partidge-coloured, as well as strains of turkeys, pheasants, quails, duck, ostriches and other poultry commonly bred in commercial quantities.


As used herein, the term “egg” refers to a fertilized ovum that has been laid by a bird. Typically, avian eggs consist of a hard, oval outer eggshell, the “egg white” or albumen, the egg yolk, and various thin membranes. Furthermore, “in ovo” refers to in an egg.


As used herein, the term “non-cellular site” refers a part of the egg other than the embryo.


The terms “reduces”, “reduction” or variations thereof as used herein refers to a measurable decrease in the amount of a target RNA and/or target protein in the egg when compared to an egg from the same species of avian, more preferably strain or breed of avian, and even more preferably the same bird, that has not been administered with a nucleic acid as defined herein. The term also refers to a measurable reduction in the activity of a target protein. Preferably a reduction in the level of a target RNA and/or target protein is at least about 10%. More preferably, the reduction is at least about 20%, 30%, 40%, 50%, 60%, 80%, 90% and even more preferably, about 100%.


As used herein, the phrase “the nucleic acid molecule results in a reduction” or variations thereof refers to the presence of the nucleic acid molecule in the egg inducing degradation of homologous RNAs in the egg by the process known in the art as “RNA interference” or “gene silencing”. Furthermore, the nucleic acid molecule directly results in the reduction, and is not transcribed in ovo to produce the desired effect.


The “at least one RNA molecule” can be any type of RNA present in, and/or produced by, an avian egg. Examples include, but are not limited to, mRNA, snRNA, microRNA and tRNA.


A “variant” of a nucleic acid molecule of the invention includes molecules of varying sizes of, and/or with one or more different nucleotides, but which are still capable of being used to silence the target gene. For example, variants may comprise additional nucleotides (such as 1, 2, 3, 4, or more), or less nucleotides. Furthermore, a few nucleotides may be substituted without influencing the ability of the nucleic acid to silence the target gene. In an embodiment, the variant includes additional 5′ and/or 3′ nucleotides which are homologous to the corresponding target RNA molecule and/or which enhance the stability of the nucleic acid molecule. In another embodiment, the nucleic acid molecules have no more than 4, more preferably no more than 3, more preferably no more than 2, and even more preferably no more than 1, nucleotide differences when compared to the sequences provided herein. In a further embodiment, the nucleic acid molecules have no more than 2, and more preferably no more than 1, internal additional and/or deletional nucleotides when compared to the sequences provided herein. In an embodiment, a nucleic acid of the invention has one, preferably two, additional non-target nucleotides at the 5′ and/or 3′ end, for example an additional UU at the 3′ end. Such additions can increase the half-life of the molecule in ovo.


By an “isolated nucleic acid molecule”, we mean a nucleic acid molecule which is at least partially separated from the nucleic acid molecule with which it is associated or linked in its native state. Preferably, the isolated nucleic acid molecule is at least 60% free, preferably at least 75% free, and most preferably at least 90% free from other components with which they are naturally associated. Furthermore, the term “polynucleotide” is used interchangeably herein with the term “nucleic acid”.


The term “exogenous” in the context of a nucleic acid molecule refers to the nucleic acid molecule when present in a cell, or in a cell-free expression system, in an altered amount. Preferably, the cell is a cell that does not naturally comprise the nucleic acid molecule. However, the cell may be a cell which comprises an exogenous nucleic acid molecule resulting in an increased amount of the nucleic acid molecule. An exogenous nucleic acid molecule of the invention includes nucleic acid molecules which have not been separated from other components of the recombinant cell, or cell-free expression system, in which it is present, and nucleic acid molecules produced in such cells or cell-free systems which are subsequently purified away from at least some other components.


Sex Determination

The present invention relates to the modulation of the sex of avians in ovo. Examples of genes which can be targeted to modulate avian sex include, but are not necessarily limited to, the DMRT1 gene, the ASW (WPKCI) gene, the R-spondin gene, the Fox9 gene and the β-catenin gene.


In a preferred embodiment, the nucleic acid molecule reduces the level of a protein encoded by a DMRT1 gene. DMRT1 was the first molecule implicated in sex determination that shows sequence conservation between phyla. The avian homologue of DMRT1 is found on the Z (sex) chromosome of chickens and is differentially expressed in the genital ridges of male and female chicken embryos (Raymond et al., 1999; Smith et al., 1999). DMRT1 is one of the few genes thus far implicated in mammalian sex determination that appears to have a strictly gonadal pattern of expression (Raymond et al., 1999).


Examples of nucleic acid molecules that can be used to reduce the level of chicken DMRT1 protein, and mRNA encoding therefor, include, but are not limited to, nucleic acids comprising one or more of the sequence of nucleotides provided in Table 1 (SEQ ID NO's 11 to 1644), or a variant of any one or more thereof, with the exception that the nucleic acid molecule does not comprise a sequence selected from:


CCAGUUGUCAAGAAGAGCA (SEQ ID NO:254)
GGAUGCUCAUUCAGGACAU (SEQ ID NO:369)
CCCUGUAUCCUUACUAUAA (SEQ ID NO:474)
GCCACUGAGUCUUCCUCAA (SEQ ID NO:530)
CCAGCAACAUACAUGUCAA (SEQ ID NO:605)
CCUGCGUCACACAGAUACU (SEQ ID NO:747)

GGAGUAGUUGUACAGGUUG (SEQ ID NO:3432) (reverse complement of SEQ ID NO:493)


GACUGGCUUGACAUGUAUG (SEQ ID NO:3433) (reverse complement of SEQ ID NO:612)


AUGGCGGUUCUCCAUCCCU (SEQ ID NO:3434) (reverse complement of SEQ ID NO:1520), or a variant of any one thereof.


In a particularly preferred embodiment, the nucleic acid molecule that can be used to reduce the level of chicken DMRT1 protein comprises a sequence selected from; GAGCCAGUUGUCAAGAAGA (SEQ ID NO:251), GACUGCCAGUGCAAGAAGU (SEQ ID NO:116), CUGUAUCCUUACUAUAACA (SEQ ID NO:476), and CUCCCAGCAACAUACAUGU (SEQ ID NO:602), or a variant of any one thereof. More preferably, the nucleic acid molecule that can be used to reduce the level of chicken DMRT1 protein comprises the sequence, GAGCCAGUUGUCAAGAAGA (SEQ ID NO:251), or a variant thereof such as GAGCCAGUUGUCAAGAAGAUU (SEQ ID NO:3431).


A further example of a gene that can be targeted to modify sex is the WPKCI gene. The avian gene WPKCI has been shown to be conserved widely on the avian W chromosome and expressed actively in the female chicken embryo before the onset of gonadal differentiation. It is suggested that WPKCI may play a role in the differentiation of the female gonad by interfering with the function of PKCI or by exhibiting its unique function in the nucleus (Hori et al., 2000). This gene has also been identified as ASW (avian sex-specific W-linked) (O'Neill et al., 2000).


Examples of nucleic acid molecules that can be used to reduce the level of chicken ASW (WPKCI) protein, and mRNA encoding therefor, include, but are not limited to, nucleic acids comprising one or more of the sequence of nucleotides provided in Table 2 (SEQ ID NO's 1645 to 2209), or a variant of any one or more thereof.


In yet another example of a gene that can be targeted to modify sex is the r-spondin gene. Examples of nucleic acid molecules that can be used to reduce the level of chicken r-spondin protein, and mRNA encoding therefor, include, but are not limited to, nucleic acids comprising one or more of the sequence of nucleotides provided in Table 3 (SEQ ID NO's 2210 to 3430), or a variant of any one or more thereof.


Gene Silencing

The terms “RNA interference”, “RNAi” or “gene silencing” refers generally to a process in which a double-stranded RNA (dsRNA) molecule reduces the expression of a nucleic acid sequence with which the double-stranded RNA molecule shares substantial or total homology. However, it has more recently been shown that gene silencing can be achieved using non-RNA double stranded molecules (see, for example, US 20070004667).


RNA interference (RNAi) is particularly useful for specifically inhibiting the production of a particular RNA and/or protein. Although not wishing to be limited by theory, Waterhouse et al. (1998) have provided a model for the mechanism by which dsRNA (duplex RNA) can be used to reduce protein production. This technology relies on the presence of dsRNA molecules that contain a sequence that is essentially identical to the mRNA of the gene of interest or part thereof, in this case an mRNA encoding a polypeptide of interest. Conveniently, the dsRNA can be produced from a single promoter in a recombinant vector or host cell, where the sense and anti-sense sequences are flanked by an unrelated sequence which enables the sense and anti-sense sequences to hybridize to form the dsRNA molecule with the unrelated sequence forming a loop structure. The design and production of suitable dsRNA molecules for the present invention is well within the capacity of a person skilled in the art, particularly considering Waterhouse et al. (1998), Smith et al. (2000), WO 99/32619, WO 99/53050, WO 99/49029 and WO 01/34815.


The present invention includes nucleic acid molecules comprising and/or encoding double-stranded regions for gene silencing. The nucleic acid molecules are typically RNA but may comprise DNA, chemically-modified nucleotides and non-nucleotides.









TABLE 1







DsRNA molecules targeting mRNA encoding chicken


DMRT1.








SEQ ID NO
Sequence 5′-3′











11
CCGGCGGCGGGCAAGAAGC





12
CGGCGGCGGGCAAGAAGCU





13
GGCGGCGGGCAAGAAGCUG





14
GCGGCGGGCAAGAAGCUGC





15
CGGCGGGCAAGAAGCUGCC





16
GGCGGGCAAGAAGCUGCCG





17
GCGGGCAAGAAGCUGCCGC





18
CGGGCAAGAAGCUGCCGCG





19
GGGCAAGAAGCUGCCGCGU





20
GGCAAGAAGCUGCCGCGUC





21
GCAAGAAGCUGCCGCGUCU





22
CAAGAAGCUGCCGCGUCUG





23
AAGAAGCUGCCGCGUCUGC





24
AGAAGCUGCCGCGUCUGCC





25
GAAGCUGCCGCGUCUGCCC





26
AAGCUGCCGCGUCUGCCCA





27
AGCUGCCGCGUCUGCCCAA





28
GCUGCCGCGUCUGCCCAAG





29
CUGCCGCGUCUGCCCAAGU





30
UGCCGCGUCUGCCCAAGUG





31
GCCGCGUCUGCCCAAGUGU





32
CCGCGUCUGCCCAAGUGUG





33
CGCGUCUGCCCAAGUGUGC





34
GCGUCUGCCCAAGUGUGCC





35
CGUCUGCCCAAGUGUGCCC





36
GUCUGCCCAAGUGUGCCCG





37
UCUGCCCAAGUGUGCCCGC





38
CUGCCCAAGUGUGCCCGCU





39
UGCCCAAGUGUGCCCGCUG





40
GCCCAAGUGUGCCCGCUGC





41
CCCAAGUGUGCCCGCUGCC





42
CCAAGUGUGCCCGCUGCCG





43
CAAGUGUGCCCGCUGCCGC





44
AAGUGUGCCCGCUGCCGCA





45
AGUGUGCCCGCUGCCGCAA





46
GUGUGCCCGCUGCCGCAAC





47
UGUGCCCGCUGCCGCAACC





48
GUGCCCGCUGCCGCAACCA





49
UGCCCGCUGCCGCAACCAC





50
GCCCGCUGCCGCAACCACG





51
CCCGCUGCCGCAACCACGG





52
CCGCUGCCGCAACCACGGC





53
CGCUGCCGCAACCACGGCU





54
GCUGCCGCAACCACGGCUA





55
CUGCCGCAACCACGGCUAC





56
UGCCGCAACCACGGCUACU





57
GCCGCAACCACGGCUACUC





58
CCGCAACCACGGCUACUCC





59
CGCAACCACGGCUACUCCU





60
GCAACCACGGCUACUCCUC





61
CAACCACGGCUACUCCUCG





62
AACCACGGCUACUCCUCGC





63
ACCACGGCUACUCCUCGCC





64
CCACGGCUACUCCUCGCCG





65
CACGGCUACUCCUCGCCGC





66
ACGGCUACUCCUCGCCGCU





67
CGGCUACUCCUCGCCGCUG





68
GGCUACUCCUCGCCGCUGA





69
GCUACUCCUCGCCGCUGAA





70
CUACUCCUCGCCGCUGAAG





71
UACUCCUCGCCGCUGAAGG





72
ACUCCUCGCCGCUGAAGGG





73
CUCCUCGCCGCUGAAGGGG





74
UCCUCGCCGCUGAAGGGGC





75
CCUCGCCGCUGAAGGGGCA





76
CUCGCCGCUGAAGGGGCAC





77
UCGCCGCUGAAGGGGCACA





78
CGCCGCUGAAGGGGCACAA





79
GCCGCUGAAGGGGCACAAG





80
CCGCUGAAGGGGCACAAGC





81
CGCUGAAGGGGCACAAGCG





82
GCUGAAGGGGCACAAGCGG





83
CUGAAGGGGCACAAGCGGU





84
UGAAGGGGCACAAGCGGUU





85
GAAGGGGCACAAGCGGUUC





86
AAGGGGCACAAGCGGUUCU





87
AGGGGCACAAGCGGUUCUG





88
GGGGCACAAGCGGUUCUGC





89
GGGCACAAGCGGUUCUGCA





90
GGCACAAGCGGUUCUGCAU





91
GCACAAGCGGUUCUGCAUG





92
CACAAGCGGUUCUGCAUGU





93
ACAAGCGGUUCUGCAUGUG





94
CAAGCGGUUCUGCAUGUGG





95
AAGCGGUUCUGCAUGUGGC





96
AGCGGUUCUGCAUGUGGCG





97
GCGGUUCUGCAUGUGGCGG





98
CGGUUCUGCAUGUGGCGGG





99
GGUUCUGCAUGUGGCGGGA





100
GUUCUGCAUGUGGCGGGAC





101
UUCUGCAUGUGGCGGGACU





102
UCUGCAUGUGGCGGGACUG





103
CUGCAUGUGGCGGGACUGC





104
UGCAUGUGGCGGGACUGCC





105
GCAUGUGGCGGGACUGCCA





106
CAUGUGGCGGGACUGCCAG





107
AUGUGGCGGGACUGCCAGU





108
UGUGGCGGGACUGCCAGUG





109
GUGGCGGGACUGCCAGUGC





110
UGGCGGGACUGCCAGUGCA





111
GGCGGGACUGCCAGUGCAA





112
GCGGGACUGCCAGUGCAAG





113
CGGGACUGCCAGUGCAAGA





114
GGGACUGCCAGUGCAAGAA





115
GGACUGCCAGUGCAAGAAG





116
GACUGCCAGUGCAAGAAGU





117
ACUGCCAGUGCAAGAAGUG





118
CUGCCAGUGCAAGAAGUGC





119
UGCCAGUGCAAGAAGUGCA





120
GCCAGUGCAAGAAGUGCAG





121
CCAGUGCAAGAAGUGCAGC





122
CAGUGCAAGAAGUGCAGCC





123
AGUGCAAGAAGUGCAGCCU





124
GUGCAAGAAGUGCAGCCUG





125
UGCAAGAAGUGCAGCCUGA





126
GCAAGAAGUGCAGCCUGAU





127
CAAGAAGUGCAGCCUGAUC





128
AAGAAGUGCAGCCUGAUCG





129
AGAAGUGCAGCCUGAUCGC





130
GAAGUGCAGCCUGAUCGCC





131
AAGUGCAGCCUGAUCGCCG





132
AGUGCAGCCUGAUCGCCGA





133
GUGCAGCCUGAUCGCCGAG





134
UGCAGCCUGAUCGCCGAGC





135
GCAGCCUGAUCGCCGAGCG





136
CAGCCUGAUCGCCGAGCGG





137
AGCCUGAUCGCCGAGCGGC





138
GCCUGAUCGCCGAGCGGCA





139
CCUGAUCGCCGAGCGGCAG





140
CUGAUCGCCGAGCGGCAGC





141
UGAUCGCCGAGCGGCAGCG





142
GAUCGCCGAGCGGCAGCGG





143
AUCGCCGAGCGGCAGCGGG





144
UCGCCGAGCGGCAGCGGGU





145
CGCCGAGCGGCAGCGGGUG





146
GCCGAGCGGCAGCGGGUGA





147
CCGAGCGGCAGCGGGUGAU





148
CGAGCGGCAGCGGGUGAUG





149
GAGCGGCAGCGGGUGAUGG





150
AGCGGCAGCGGGUGAUGGC





151
GCGGCAGCGGGUGAUGGCC





152
CGGCAGCGGGUGAUGGCCG





153
GGCAGCGGGUGAUGGCCGU





154
GCAGCGGGUGAUGGCCGUG





155
CAGCGGGUGAUGGCCGUGC





156
AGCGGGUGAUGGCCGUGCA





157
GCGGGUGAUGGCCGUGCAG





158
CGGGUGAUGGCCGUGCAGG





159
GGGUGAUGGCCGUGCAGGU





160
GGUGAUGGCCGUGCAGGUU





161
GUGAUGGCCGUGCAGGUUG





162
UGAUGGCCGUGCAGGUUGC





163
GAUGGCCGUGCAGGUUGCA





164
AUGGCCGUGCAGGUUGCAC





165
UGGCCGUGCAGGUUGCACU





166
GGCCGUGCAGGUUGCACUG





167
GCCGUGCAGGUUGCACUGA





168
CCGUGCAGGUUGCACUGAG





169
CGUGCAGGUUGCACUGAGG





170
GUGCAGGUUGCACUGAGGA





171
UGCAGGUUGCACUGAGGAG





172
GCAGGUUGCACUGAGGAGG





173
CAGGUUGCACUGAGGAGGC





174
AGGUUGCACUGAGGAGGCA





175
GGUUGCACUGAGGAGGCAG





176
GUUGCACUGAGGAGGCAGC





177
UUGCACUGAGGAGGCAGCA





178
UGCACUGAGGAGGCAGCAA





179
GCACUGAGGAGGCAGCAAG





180
CACUGAGGAGGCAGCAAGC





181
ACUGAGGAGGCAGCAAGCC





182
CUGAGGAGGCAGCAAGCCC





183
UGAGGAGGCAGCAAGCCCA





184
GAGGAGGCAGCAAGCCCAG





185
AGGAGGCAGCAAGCCCAGG





186
GGAGGCAGCAAGCCCAGGA





187
GAGGCAGCAAGCCCAGGAA





188
AGGCAGCAAGCCCAGGAAG





189
GGCAGCAAGCCCAGGAAGA





190
GCAGCAAGCCCAGGAAGAG





191
CAGCAAGCCCAGGAAGAGG





192
AGCAAGCCCAGGAAGAGGA





193
GCAAGCCCAGGAAGAGGAG





194
CAAGCCCAGGAAGAGGAGC





195
AAGCCCAGGAAGAGGAGCU





196
AGCCCAGGAAGAGGAGCUG





197
GCCCAGGAAGAGGAGCUGG





198
CCCAGGAAGAGGAGCUGGG





199
CCAGGAAGAGGAGCUGGGG





200
CAGGAAGAGGAGCUGGGGA





201
AGGAAGAGGAGCUGGGGAU





202
GGAAGAGGAGCUGGGGAUC





203
GAAGAGGAGCUGGGGAUCA





204
AAGAGGAGCUGGGGAUCAG





205
AGAGGAGCUGGGGAUCAGC





206
GAGGAGCUGGGGAUCAGCC





207
AGGAGCUGGGGAUCAGCCA





208
GGAGCUGGGGAUCAGCCAC





209
GAGCUGGGGAUCAGCCACC





210
AGCUGGGGAUCAGCCACCC





211
GCUGGGGAUCAGCCACCCU





212
CUGGGGAUCAGCCACCCUG





213
UGGGGAUCAGCCACCCUGU





214
GGGGAUCAGCCACCCUGUA





215
GGGAUCAGCCACCCUGUAC





216
GGAUCAGCCACCCUGUACC





217
GAUCAGCCACCCUGUACCC





218
AUCAGCCACCCUGUACCCC





219
UCAGCCACCCUGUACCCCU





220
CAGCCACCCUGUACCCCUG





221
AGCCACCCUGUACCCCUGC





222
GCCACCCUGUACCCCUGCC





223
CCACCCUGUACCCCUGCCC





224
CACCCUGUACCCCUGCCCA





225
ACCCUGUACCCCUGCCCAG





226
CCCUGUACCCCUGCCCAGU





227
CCUGUACCCCUGCCCAGUG





228
CUGUACCCCUGCCCAGUGC





229
UGUACCCCUGCCCAGUGCC





230
GUACCCCUGCCCAGUGCCC





231
UACCCCUGCCCAGUGCCCC





232
ACCCCUGCCCAGUGCCCCU





233
CCCCUGCCCAGUGCCCCUG





234
CCCUGCCCAGUGCCCCUGA





235
CCUGCCCAGUGCCCCUGAG





236
CUGCCCAGUGCCCCUGAGC





237
UGCCCAGUGCCCCUGAGCC





238
GCCCAGUGCCCCUGAGCCA





239
CCCAGUGCCCCUGAGCCAG





240
CCAGUGCCCCUGAGCCAGU





241
CAGUGCCCCUGAGCCAGUU





242
AGUGCCCCUGAGCCAGUUG





243
GUGCCCCUGAGCCAGUUGU





244
UGCCCCUGAGCCAGUUGUC





245
GCCCCUGAGCCAGUUGUCA





246
CCCCUGAGCCAGUUGUCAA





247
CCCUGAGCCAGUUGUCAAG





248
CCUGAGCCAGUUGUCAAGA





249
CUGAGCCAGUUGUCAAGAA





250
UGAGCCAGUUGUCAAGAAG





251
GAGCCAGUUGUCAAGAAGA





252
AGCCAGUUGUCAAGAAGAG





253
GCCAGUUGUCAAGAAGAGC





254
CCAGUUGUCAAGAAGAGCA





255
CAGUUGUCAAGAAGAGCAG





256
AGUUGUCAAGAAGAGCAGC





257
GUUGUCAAGAAGAGCAGCA





258
UUGUCAAGAAGAGCAGCAG





259
UGUCAAGAAGAGCAGCAGC





260
GUCAAGAAGAGCAGCAGCA





261
UCAAGAAGAGCAGCAGCAG





262
CAAGAAGAGCAGCAGCAGC





263
AAGAAGAGCAGCAGCAGCA





264
AGAAGAGCAGCAGCAGCAG





265
GAAGAGCAGCAGCAGCAGC





266
AAGAGCAGCAGCAGCAGCU





267
AGAGCAGCAGCAGCAGCUC





268
GAGCAGCAGCAGCAGCUCC





269
AGCAGCAGCAGCAGCUCCU





270
GCAGCAGCAGCAGCUCCUG





271
CAGCAGCAGCAGCUCCUGU





272
AGCAGCAGCAGCUCCUGUC





273
GCAGCAGCAGCUCCUGUCU





274
CAGCAGCAGCUCCUGUCUC





275
AGCAGCAGCUCCUGUCUCC





276
GCAGCAGCUCCUGUCUCCU





277
CAGCAGCUCCUGUCUCCUG





278
AGCAGCUCCUGUCUCCUGC





279
GCAGCUCCUGUCUCCUGCA





280
CAGCUCCUGUCUCCUGCAG





281
AGCUCCUGUCUCCUGCAGG





282
GCUCCUGUCUCCUGCAGGA





283
CUCCUGUCUCCUGCAGGAC





284
UCCUGUCUCCUGCAGGACA





285
CCUGUCUCCUGCAGGACAG





286
CUGUCUCCUGCAGGACAGC





287
UGUCUCCUGCAGGACAGCA





288
GUCUCCUGCAGGACAGCAG





289
UCUCCUGCAGGACAGCAGC





290
CUCCUGCAGGACAGCAGCA





291
UCCUGCAGGACAGCAGCAG





292
CCUGCAGGACAGCAGCAGC





293
CUGCAGGACAGCAGCAGCC





294
UGCAGGACAGCAGCAGCCC





295
GCAGGACAGCAGCAGCCCU





296
CAGGACAGCAGCAGCCCUG





297
AGGACAGCAGCAGCCCUGC





298
GGACAGCAGCAGCCCUGCU





299
GACAGCAGCAGCCCUGCUC





300
ACAGCAGCAGCCCUGCUCA





301
CAGCAGCAGCCCUGCUCAC





302
AGCAGCAGCCCUGCUCACU





303
GCAGCAGCCCUGCUCACUC





304
CAGCAGCCCUGCUCACUCC





305
AGCAGCCCUGCUCACUCCA





306
GCAGCCCUGCUCACUCCAC





307
CAGCCCUGCUCACUCCACG





308
AGCCCUGCUCACUCCACGA





309
GCCCUGCUCACUCCACGAG





310
CCCUGCUCACUCCACGAGC





311
CCUGCUCACUCCACGAGCA





312
CUGCUCACUCCACGAGCAC





313
UGCUCACUCCACGAGCACG





314
GCUCACUCCACGAGCACGG





315
CUCACUCCACGAGCACGGU





316
UCACUCCACGAGCACGGUG





317
CACUCCACGAGCACGGUGG





318
ACUCCACGAGCACGGUGGC





319
CUCCACGAGCACGGUGGCA





320
UCCACGAGCACGGUGGCAG





321
CCACGAGCACGGUGGCAGC





322
CACGAGCACGGUGGCAGCA





323
ACGAGCACGGUGGCAGCAG





324
CGAGCACGGUGGCAGCAGC





325
GAGCACGGUGGCAGCAGCA





326
AGCACGGUGGCAGCAGCAG





327
GCACGGUGGCAGCAGCAGC





328
CACGGUGGCAGCAGCAGCA





329
ACGGUGGCAGCAGCAGCAG





330
CGGUGGCAGCAGCAGCAGC





331
GGUGGCAGCAGCAGCAGCG





332
GUGGCAGCAGCAGCAGCGA





333
UGGCAGCAGCAGCAGCGAG





334
GGCAGCAGCAGCAGCGAGC





335
GCAGCAGCAGCAGCGAGCG





336
CAGCAGCAGCAGCGAGCGC





337
AGCAGCAGCAGCGAGCGCA





338
GCAGCAGCAGCGAGCGCAC





339
CAGCAGCAGCGAGCGCACC





340
AGCAGCAGCGAGCGCACCA





341
GCAGCAGCGAGCGCACCAC





342
CAGCAGCGAGCGCACCACC





343
AGCAGCGAGCGCACCACCA





344
GCAGCGAGCGCACCACCAG





345
CAGCGAGCGCACCACCAGA





346
AGCGAGCGCACCACCAGAG





347
GCGAGCGCACCACCAGAGG





348
CGAGCGCACCACCAGAGGG





349
GAGCGCACCACCAGAGGGA





350
AGCGCACCACCAGAGGGAC





351
GCGCACCACCAGAGGGACG





352
CGCACCACCAGAGGGACGG





353
GCACCACCAGAGGGACGGA





354
CACCACCAGAGGGACGGAU





355
ACCACCAGAGGGACGGAUG





356
CCACCAGAGGGACGGAUGC





357
CACCAGAGGGACGGAUGCU





358
ACCAGAGGGACGGAUGCUC





359
CCAGAGGGACGGAUGCUCA





360
CAGAGGGACGGAUGCUCAU





361
AGAGGGACGGAUGCUCAUU





362
GAGGGACGGAUGCUCAUUC





363
AGGGACGGAUGCUCAUUCA





364
GGGACGGAUGCUCAUUCAG





365
GGACGGAUGCUCAUUCAGG





366
GACGGAUGCUCAUUCAGGA





367
ACGGAUGCUCAUUCAGGAC





368
CGGAUGCUCAUUCAGGACA





369
GGAUGCUCAUUCAGGACAU





370
GAUGCUCAUUCAGGACAUC





371
AUGCUCAUUCAGGACAUCC





372
UGCUCAUUCAGGACAUCCC





373
GCUCAUUCAGGACAUCCCU





374
CUCAUUCAGGACAUCCCUU





375
UCAUUCAGGACAUCCCUUC





376
CAUUCAGGACAUCCCUUCC





377
AUUCAGGACAUCCCUUCCA





378
UUCAGGACAUCCCUUCCAU





379
UCAGGACAUCCCUUCCAUC





380
CAGGACAUCCCUUCCAUCC





381
AGGACAUCCCUUCCAUCCC





382
GGACAUCCCUUCCAUCCCC





383
GACAUCCCUUCCAUCCCCA





384
ACAUCCCUUCCAUCCCCAG





385
CAUCCCUUCCAUCCCCAGC





386
AUCCCUUCCAUCCCCAGCA





387
UCCCUUCCAUCCCCAGCAG





388
CCCUUCCAUCCCCAGCAGA





389
CCUUCCAUCCCCAGCAGAG





390
CUUCCAUCCCCAGCAGAGG





391
UUCCAUCCCCAGCAGAGGG





392
UCCAUCCCCAGCAGAGGGC





393
CCAUCCCCAGCAGAGGGCA





394
CAUCCCCAGCAGAGGGCAC





395
AUCCCCAGCAGAGGGCACU





396
UCCCCAGCAGAGGGCACUU





397
CCCCAGCAGAGGGCACUUG





398
CCCAGCAGAGGGCACUUGG





399
CCAGCAGAGGGCACUUGGA





400
CAGCAGAGGGCACUUGGAG





401
AGCAGAGGGCACUUGGAGA





402
GCAGAGGGCACUUGGAGAG





403
CAGAGGGCACUUGGAGAGC





404
AGAGGGCACUUGGAGAGCA





405
GAGGGCACUUGGAGAGCAC





406
AGGGCACUUGGAGAGCACG





407
GGGCACUUGGAGAGCACGU





408
GGCACUUGGAGAGCACGUC





409
GCACUUGGAGAGCACGUCU





410
CACUUGGAGAGCACGUCUG





411
ACUUGGAGAGCACGUCUGA





412
CUUGGAGAGCACGUCUGAU





413
UUGGAGAGCACGUCUGAUU





414
UGGAGAGCACGUCUGAUUU





415
GGAGAGCACGUCUGAUUUG





416
GAGAGCACGUCUGAUUUGG





417
AGAGCACGUCUGAUUUGGU





418
GAGCACGUCUGAUUUGGUU





419
AGCACGUCUGAUUUGGUUG





420
GCACGUCUGAUUUGGUUGU





421
CACGUCUGAUUUGGUUGUG





422
ACGUCUGAUUUGGUUGUGG





423
CGUCUGAUUUGGUUGUGGA





424
GUCUGAUUUGGUUGUGGAC





425
UCUGAUUUGGUUGUGGACU





426
CUGAUUUGGUUGUGGACUC





427
UGAUUUGGUUGUGGACUCC





428
GAUUUGGUUGUGGACUCCA





429
AUUUGGUUGUGGACUCCAC





430
UUUGGUUGUGGACUCCACC





431
UUGGUUGUGGACUCCACCU





432
UGGUUGUGGACUCCACCUA





433
GGUUGUGGACUCCACCUAC





434
GUUGUGGACUCCACCUACU





435
UUGUGGACUCCACCUACUA





436
UGUGGACUCCACCUACUAC





437
GUGGACUCCACCUACUACA





438
UGGACUCCACCUACUACAG





439
GGACUCCACCUACUACAGC





440
GACUCCACCUACUACAGCA





441
ACUCCACCUACUACAGCAG





442
CUCCACCUACUACAGCAGU





443
UCCACCUACUACAGCAGUU





444
CCACCUACUACAGCAGUUU





445
CACCUACUACAGCAGUUUU





446
ACCUACUACAGCAGUUUUU





447
CCUACUACAGCAGUUUUUA





448
CUACUACAGCAGUUUUUAC





449
UACUACAGCAGUUUUUACC





450
ACUACAGCAGUUUUUACCA





451
CUACAGCAGUUUUUACCAG





452
UACAGCAGUUUUUACCAGC





453
ACAGCAGUUUUUACCAGCC





454
CAGCAGUUUUUACCAGCCA





455
AGCAGUUUUUACCAGCCAU





456
GCAGUUUUUACCAGCCAUC





457
CAGUUUUUACCAGCCAUCC





458
AGUUUUUACCAGCCAUCCC





459
GUUUUUACCAGCCAUCCCU





460
UUUUUACCAGCCAUCCCUG





461
UUUUACCAGCCAUCCCUGU





462
UUUACCAGCCAUCCCUGUA





463
UUACCAGCCAUCCCUGUAU





464
UACCAGCCAUCCCUGUAUC





465
ACCAGCCAUCCCUGUAUCC





466
CCAGCCAUCCCUGUAUCCU





467
CAGCCAUCCCUGUAUCCUU





468
AGCCAUCCCUGUAUCCUUA





469
GCCAUCCCUGUAUCCUUAC





470
CCAUCCCUGUAUCCUUACU





471
CAUCCCUGUAUCCUUACUA





472
AUCCCUGUAUCCUUACUAU





473
UCCCUGUAUCCUUACUAUA





474
CCCUGUAUCCUUACUAUAA





475
CCUGUAUCCUUACUAUAAC





476
CUGUAUCCUUACUAUAACA





477
UGUAUCCUUACUAUAACAA





478
GUAUCCUUACUAUAACAAC





479
UAUCCUUACUAUAACAACC





480
AUCCUUACUAUAACAACCU





481
UCCUUACUAUAACAACCUG





482
CCUUACUAUAACAACCUGU





483
CUUACUAUAACAACCUGUA





484
UUACUAUAACAACCUGUAC





485
UACUAUAACAACCUGUACA





486
ACUAUAACAACCUGUACAA





487
CUAUAACAACCUGUACAAC





488
UAUAACAACCUGUACAACU





489
AUAACAACCUGUACAACUA





490
UAACAACCUGUACAACUAC





491
AACAACCUGUACAACUACU





492
ACAACCUGUACAACUACUC





493
CAACCUGUACAACUACUCC





494
AACCUGUACAACUACUCCC





495
ACCUGUACAACUACUCCCA





496
CCUGUACAACUACUCCCAG





497
CUGUACAACUACUCCCAGU





498
UGUACAACUACUCCCAGUA





499
GUACAACUACUCCCAGUAC





500
UACAACUACUCCCAGUACC





501
ACAACUACUCCCAGUACCA





502
CAACUACUCCCAGUACCAA





503
AACUACUCCCAGUACCAAA





504
ACUACUCCCAGUACCAAAU





505
CUACUCCCAGUACCAAAUG





506
UACUCCCAGUACCAAAUGG





507
ACUCCCAGUACCAAAUGGC





508
CUCCCAGUACCAAAUGGCA





509
UCCCAGUACCAAAUGGCAG





510
CCCAGUACCAAAUGGCAGU





511
CCAGUACCAAAUGGCAGUG





512
CAGUACCAAAUGGCAGUGG





513
AGUACCAAAUGGCAGUGGC





514
GUACCAAAUGGCAGUGGCC





515
UACCAAAUGGCAGUGGCCA





516
ACCAAAUGGCAGUGGCCAC





517
CCAAAUGGCAGUGGCCACU





518
CAAAUGGCAGUGGCCACUG





519
AAAUGGCAGUGGCCACUGA





520
AAUGGCAGUGGCCACUGAG





521
AUGGCAGUGGCCACUGAGU





522
UGGCAGUGGCCACUGAGUC





523
GGCAGUGGCCACUGAGUCU





524
GCAGUGGCCACUGAGUCUU





525
CAGUGGCCACUGAGUCUUC





526
AGUGGCCACUGAGUCUUCC





527
GUGGCCACUGAGUCUUCCU





528
UGGCCACUGAGUCUUCCUC





529
GGCCACUGAGUCUUCCUCA





530
GCCACUGAGUCUUCCUCAA





531
CCACUGAGUCUUCCUCAAG





532
CACUGAGUCUUCCUCAAGU





533
ACUGAGUCUUCCUCAAGUG





534
CUGAGUCUUCCUCAAGUGA





535
UGAGUCUUCCUCAAGUGAG





536
GAGUCUUCCUCAAGUGAGA





537
AGUCUUCCUCAAGUGAGAC





538
GUCUUCCUCAAGUGAGACA





539
UCUUCCUCAAGUGAGACAG





540
CUUCCUCAAGUGAGACAGG





541
UUCCUCAAGUGAGACAGGG





542
UCCUCAAGUGAGACAGGGG





543
CCUCAAGUGAGACAGGGGG





544
CUCAAGUGAGACAGGGGGU





545
UCAAGUGAGACAGGGGGUA





546
CAAGUGAGACAGGGGGUAC





547
AAGUGAGACAGGGGGUACG





548
AGUGAGACAGGGGGUACGU





549
GUGAGACAGGGGGUACGUU





550
UGAGACAGGGGGUACGUUU





551
GAGACAGGGGGUACGUUUG





552
AGACAGGGGGUACGUUUGU





553
GACAGGGGGUACGUUUGUA





554
ACAGGGGGUACGUUUGUAG





555
CAGGGGGUACGUUUGUAGG





556
AGGGGGUACGUUUGUAGGG





557
GGGGGUACGUUUGUAGGGU





558
GGGGUACGUUUGUAGGGUC





559
GGGUACGUUUGUAGGGUCA





560
GGUACGUUUGUAGGGUCAG





561
GUACGUUUGUAGGGUCAGC





562
UACGUUUGUAGGGUCAGCC





563
ACGUUUGUAGGGUCAGCCA





564
CGUUUGUAGGGUCAGCCAU





565
GUUUGUAGGGUCAGCCAUG





566
UUUGUAGGGUCAGCCAUGA





567
UUGUAGGGUCAGCCAUGAA





568
UGUAGGGUCAGCCAUGAAA





569
GUAGGGUCAGCCAUGAAAA





570
UAGGGUCAGCCAUGAAAAA





571
AGGGUCAGCCAUGAAAAAC





572
GGGUCAGCCAUGAAAAACA





573
GGUCAGCCAUGAAAAACAG





574
GUCAGCCAUGAAAAACAGC





575
UCAGCCAUGAAAAACAGCC





576
CAGCCAUGAAAAACAGCCU





577
AGCCAUGAAAAACAGCCUU





578
GCCAUGAAAAACAGCCUUC





579
CCAUGAAAAACAGCCUUCG





580
CAUGAAAAACAGCCUUCGA





581
AUGAAAAACAGCCUUCGAA





582
UGAAAAACAGCCUUCGAAG





583
GAAAAACAGCCUUCGAAGC





584
AAAAACAGCCUUCGAAGCC





585
AAAACAGCCUUCGAAGCCU





586
AAACAGCCUUCGAAGCCUC





587
AACAGCCUUCGAAGCCUCC





588
ACAGCCUUCGAAGCCUCCC





589
CAGCCUUCGAAGCCUCCCA





590
AGCCUUCGAAGCCUCCCAG





591
GCCUUCGAAGCCUCCCAGC





592
CCUUCGAAGCCUCCCAGCA





593
CUUCGAAGCCUCCCAGCAA





594
UUCGAAGCCUCCCAGCAAC





595
UCGAAGCCUCCCAGCAACA





596
CGAAGCCUCCCAGCAACAU





597
GAAGCCUCCCAGCAACAUA





598
AAGCCUCCCAGCAACAUAC





599
AGCCUCCCAGCAACAUACA





600
GCCUCCCAGCAACAUACAU





601
CCUCCCAGCAACAUACAUG





602
CUCCCAGCAACAUACAUGU





603
UCCCAGCAACAUACAUGUC





604
CCCAGCAACAUACAUGUCA





605
CCAGCAACAUACAUGUCAA





606
CAGCAACAUACAUGUCAAG





607
AGCAACAUACAUGUCAAGC





608
GCAACAUACAUGUCAAGCC





609
CAACAUACAUGUCAAGCCA





610
AACAUACAUGUCAAGCCAG





611
ACAUACAUGUCAAGCCAGU





612
CAUACAUGUCAAGCCAGUC





613
AUACAUGUCAAGCCAGUCA





614
UACAUGUCAAGCCAGUCAG





615
ACAUGUCAAGCCAGUCAGG





616
CAUGUCAAGCCAGUCAGGA





617
AUGUCAAGCCAGUCAGGAA





618
UGUCAAGCCAGUCAGGAAA





619
GUCAAGCCAGUCAGGAAAA





620
UCAAGCCAGUCAGGAAAAC





621
CAAGCCAGUCAGGAAAACA





622
AAGCCAGUCAGGAAAACAG





623
AGCCAGUCAGGAAAACAGU





624
GCCAGUCAGGAAAACAGUG





625
CCAGUCAGGAAAACAGUGG





626
CAGUCAGGAAAACAGUGGC





627
AGUCAGGAAAACAGUGGCA





628
GUCAGGAAAACAGUGGCAG





629
UCAGGAAAACAGUGGCAGA





630
CAGGAAAACAGUGGCAGAU





631
AGGAAAACAGUGGCAGAUG





632
GGAAAACAGUGGCAGAUGA





633
GAAAACAGUGGCAGAUGAA





634
AAAACAGUGGCAGAUGAAG





635
AAACAGUGGCAGAUGAAGG





636
AACAGUGGCAGAUGAAGGG





637
ACAGUGGCAGAUGAAGGGA





638
CAGUGGCAGAUGAAGGGAA





639
AGUGGCAGAUGAAGGGAAU





640
GUGGCAGAUGAAGGGAAUG





641
UGGCAGAUGAAGGGAAUGG





642
GGCAGAUGAAGGGAAUGGA





643
GCAGAUGAAGGGAAUGGAG





644
CAGAUGAAGGGAAUGGAGA





645
AGAUGAAGGGAAUGGAGAA





646
GAUGAAGGGAAUGGAGAAC





647
AUGAAGGGAAUGGAGAACC





648
UGAAGGGAAUGGAGAACCG





649
GAAGGGAAUGGAGAACCGC





650
AAGGGAAUGGAGAACCGCC





651
AGGGAAUGGAGAACCGCCA





652
GGGAAUGGAGAACCGCCAU





653
GGAAUGGAGAACCGCCAUG





654
GAAUGGAGAACCGCCAUGC





655
AAUGGAGAACCGCCAUGCC





656
AUGGAGAACCGCCAUGCCA





657
UGGAGAACCGCCAUGCCAU





658
GGAGAACCGCCAUGCCAUG





659
GAGAACCGCCAUGCCAUGA





660
AGAACCGCCAUGCCAUGAG





661
GAACCGCCAUGCCAUGAGC





662
AACCGCCAUGCCAUGAGCU





663
ACCGCCAUGCCAUGAGCUC





664
CCGCCAUGCCAUGAGCUCC





665
CGCCAUGCCAUGAGCUCCC





666
GCCAUGCCAUGAGCUCCCA





667
CCAUGCCAUGAGCUCCCAG





668
CAUGCCAUGAGCUCCCAGU





669
AUGCCAUGAGCUCCCAGUA





670
UGCCAUGAGCUCCCAGUAC





671
GCCAUGAGCUCCCAGUACC





672
CCAUGAGCUCCCAGUACCG





673
CAUGAGCUCCCAGUACCGG





674
AUGAGCUCCCAGUACCGGA





675
UGAGCUCCCAGUACCGGAU





676
GAGCUCCCAGUACCGGAUG





677
AGCUCCCAGUACCGGAUGU





678
GCUCCCAGUACCGGAUGUG





679
CUCCCAGUACCGGAUGUGC





680
UCCCAGUACCGGAUGUGCU





681
CCCAGUACCGGAUGUGCUC





682
CCAGUACCGGAUGUGCUCC





683
CAGUACCGGAUGUGCUCCU





684
AGUACCGGAUGUGCUCCUA





685
GUACCGGAUGUGCUCCUAC





686
UACCGGAUGUGCUCCUACU





687
ACCGGAUGUGCUCCUACUA





688
CCGGAUGUGCUCCUACUAC





689
CGGAUGUGCUCCUACUACC





690
GGAUGUGCUCCUACUACCC





691
GAUGUGCUCCUACUACCCG





692
AUGUGCUCCUACUACCCGC





693
UGUGCUCCUACUACCCGCC





694
GUGCUCCUACUACCCGCCC





695
UGCUCCUACUACCCGCCCA





696
GCUCCUACUACCCGCCCAC





697
CUCCUACUACCCGCCCACC





698
UCCUACUACCCGCCCACCU





699
CCUACUACCCGCCCACCUC





700
CUACUACCCGCCCACCUCA





701
UACUACCCGCCCACCUCAU





702
ACUACCCGCCCACCUCAUA





703
CUACCCGCCCACCUCAUAC





704
UACCCGCCCACCUCAUACC





705
ACCCGCCCACCUCAUACCU





706
CCCGCCCACCUCAUACCUG





707
CCGCCCACCUCAUACCUGG





708
CGCCCACCUCAUACCUGGG





709
GCCCACCUCAUACCUGGGC





710
CCCACCUCAUACCUGGGCC





711
CCACCUCAUACCUGGGCCA





712
CACCUCAUACCUGGGCCAG





713
ACCUCAUACCUGGGCCAGG





714
CCUCAUACCUGGGCCAGGG





715
CUCAUACCUGGGCCAGGGG





716
UCAUACCUGGGCCAGGGGG





717
CAUACCUGGGCCAGGGGGU





718
AUACCUGGGCCAGGGGGUU





719
UACCUGGGCCAGGGGGUUG





720
ACCUGGGCCAGGGGGUUGG





721
CCUGGGCCAGGGGGUUGGC





722
CUGGGCCAGGGGGUUGGCA





723
UGGGCCAGGGGGUUGGCAG





724
GGGCCAGGGGGUUGGCAGU





725
GGCCAGGGGGUUGGCAGUC





726
GCCAGGGGGUUGGCAGUCC





727
CCAGGGGGUUGGCAGUCCC





728
CAGGGGGUUGGCAGUCCCA





729
AGGGGGUUGGCAGUCCCAC





730
GGGGGUUGGCAGUCCCACC





731
GGGGUUGGCAGUCCCACCU





732
GGGUUGGCAGUCCCACCUG





733
GGUUGGCAGUCCCACCUGC





734
GUUGGCAGUCCCACCUGCG





735
UUGGCAGUCCCACCUGCGU





736
UGGCAGUCCCACCUGCGUC





737
GGCAGUCCCACCUGCGUCA





738
GCAGUCCCACCUGCGUCAC





739
CAGUCCCACCUGCGUCACA





740
AGUCCCACCUGCGUCACAC





741
GUCCCACCUGCGUCACACA





742
UCCCACCUGCGUCACACAG





743
CCCACCUGCGUCACACAGA





744
CCACCUGCGUCACACAGAU





745
CACCUGCGUCACACAGAUA





746
ACCUGCGUCACACAGAUAC





747
CCUGCGUCACACAGAUACU





748
CUGCGUCACACAGAUACUG





749
UGCGUCACACAGAUACUGG





750
GCGUCACACAGAUACUGGC





751
CGUCACACAGAUACUGGCC





752
GUCACACAGAUACUGGCCU





753
UCACACAGAUACUGGCCUC





754
CACACAGAUACUGGCCUCG





755
ACACAGAUACUGGCCUCGG





756
CACAGAUACUGGCCUCGGA





757
ACAGAUACUGGCCUCGGAG





758
CAGAUACUGGCCUCGGAGG





759
AGAUACUGGCCUCGGAGGA





760
GAUACUGGCCUCGGAGGAC





761
AUACUGGCCUCGGAGGACA





762
UACUGGCCUCGGAGGACAC





763
ACUGGCCUCGGAGGACACC





764
CUGGCCUCGGAGGACACCC





765
UGGCCUCGGAGGACACCCC





766
GGCCUCGGAGGACACCCCC





767
GCCUCGGAGGACACCCCCU





768
CCUCGGAGGACACCCCCUC





769
CUCGGAGGACACCCCCUCC





770
UCGGAGGACACCCCCUCCU





771
CGGAGGACACCCCCUCCUA





772
GGAGGACACCCCCUCCUAC





773
GAGGACACCCCCUCCUACU





774
AGGACACCCCCUCCUACUC





775
GGACACCCCCUCCUACUCA





776
GACACCCCCUCCUACUCAG





777
ACACCCCCUCCUACUCAGA





778
CACCCCCUCCUACUCAGAG





779
ACCCCCUCCUACUCAGAGU





780
CCCCCUCCUACUCAGAGUC





781
CCCCUCCUACUCAGAGUCG





782
CCCUCCUACUCAGAGUCGA





783
CCUCCUACUCAGAGUCGAA





784
CUCCUACUCAGAGUCGAAA





785
UCCUACUCAGAGUCGAAAG





786
CCUACUCAGAGUCGAAAGC





787
CUACUCAGAGUCGAAAGCG





788
UACUCAGAGUCGAAAGCGA





789
ACUCAGAGUCGAAAGCGAG





790
CUCAGAGUCGAAAGCGAGA





791
UCAGAGUCGAAAGCGAGAG





792
CAGAGUCGAAAGCGAGAGU





793
AGAGUCGAAAGCGAGAGUG





794
GAGUCGAAAGCGAGAGUGU





795
AGUCGAAAGCGAGAGUGUU





796
GUCGAAAGCGAGAGUGUUU





797
UCGAAAGCGAGAGUGUUUU





798
CGAAAGCGAGAGUGUUUUC





799
GAAAGCGAGAGUGUUUUCG





800
AAAGCGAGAGUGUUUUCGC





801
AAGCGAGAGUGUUUUCGCC





802
AGCGAGAGUGUUUUCGCCG





803
GCGAGAGUGUUUUCGCCGC





804
CGAGAGUGUUUUCGCCGCC





805
GAGAGUGUUUUCGCCGCCC





806
AGAGUGUUUUCGCCGCCCA





807
GAGUGUUUUCGCCGCCCAG





808
AGUGUUUUCGCCGCCCAGC





809
GUGUUUUCGCCGCCCAGCA





810
UGUUUUCGCCGCCCAGCAG





811
GUUUUCGCCGCCCAGCAGC





812
UUUUCGCCGCCCAGCAGCC





813
UUUCGCCGCCCAGCAGCCA





814
UUCGCCGCCCAGCAGCCAG





815
UCGCCGCCCAGCAGCCAGG





816
CGCCGCCCAGCAGCCAGGA





817
GCCGCCCAGCAGCCAGGAC





818
CCGCCCAGCAGCCAGGACU





819
CGCCCAGCAGCCAGGACUC





820
GCCCAGCAGCCAGGACUCG





821
CCCAGCAGCCAGGACUCGG





822
CCAGCAGCCAGGACUCGGG





823
CAGCAGCCAGGACUCGGGC





824
AGCAGCCAGGACUCGGGCC





825
GCAGCCAGGACUCGGGCCU





826
CAGCCAGGACUCGGGCCUG





827
AGCCAGGACUCGGGCCUGG





828
GCCAGGACUCGGGCCUGGG





829
CCAGGACUCGGGCCUGGGG





830
CAGGACUCGGGCCUGGGGU





831
AGGACUCGGGCCUGGGGUG





832
GGACUCGGGCCUGGGGUGC





833
GACUCGGGCCUGGGGUGCC





834
ACUCGGGCCUGGGGUGCCU





835
CUCGGGCCUGGGGUGCCUG





836
UCGGGCCUGGGGUGCCUGU





837
CGGGCCUGGGGUGCCUGUC





838
GGGCCUGGGGUGCCUGUCG





839
GGCCUGGGGUGCCUGUCGA





840
GCCUGGGGUGCCUGUCGAG





841
CCUGGGGUGCCUGUCGAGC





842
CUGGGGUGCCUGUCGAGCA





843
UGGGGUGCCUGUCGAGCAG





844
GGGGUGCCUGUCGAGCAGC





845
GGGUGCCUGUCGAGCAGCG





846
GGUGCCUGUCGAGCAGCGA





847
GUGCCUGUCGAGCAGCGAG





848
UGCCUGUCGAGCAGCGAGA





849
GCCUGUCGAGCAGCGAGAG





850
CCUGUCGAGCAGCGAGAGC





851
CUGUCGAGCAGCGAGAGCA





852
UGUCGAGCAGCGAGAGCAC





853
GUCGAGCAGCGAGAGCACC





854
UCGAGCAGCGAGAGCACCA





855
CGAGCAGCGAGAGCACCAA





856
GAGCAGCGAGAGCACCAAG





857
AGCAGCGAGAGCACCAAGG





858
GCAGCGAGAGCACCAAGGG





859
CAGCGAGAGCACCAAGGGA





860
AGCGAGAGCACCAAGGGAG





861
GCGAGAGCACCAAGGGAGA





862
CGAGAGCACCAAGGGAGAC





863
GAGAGCACCAAGGGAGACC





864
AGAGCACCAAGGGAGACCU





865
GAGCACCAAGGGAGACCUG





866
AGCACCAAGGGAGACCUGG





867
GCACCAAGGGAGACCUGGA





868
CACCAAGGGAGACCUGGAG





869
ACCAAGGGAGACCUGGAGU





870
CCAAGGGAGACCUGGAGUG





871
CAAGGGAGACCUGGAGUGC





872
AAGGGAGACCUGGAGUGCG





873
AGGGAGACCUGGAGUGCGA





874
GGGAGACCUGGAGUGCGAG





875
GGAGACCUGGAGUGCGAGC





876
GAGACCUGGAGUGCGAGCC





877
AGACCUGGAGUGCGAGCCC





878
GACCUGGAGUGCGAGCCCC





879
ACCUGGAGUGCGAGCCCCA





880
CCUGGAGUGCGAGCCCCAC





881
CUGGAGUGCGAGCCCCACC





882
UGGAGUGCGAGCCCCACCA





883
GGAGUGCGAGCCCCACCAA





884
GAGUGCGAGCCCCACCAAG





885
AGUGCGAGCCCCACCAAGA





886
GUGCGAGCCCCACCAAGAG





887
UGCGAGCCCCACCAAGAGC





888
GCGAGCCCCACCAAGAGCC





889
CGAGCCCCACCAAGAGCCC





890
GAGCCCCACCAAGAGCCCG





891
AGCCCCACCAAGAGCCCGG





892
GCCCCACCAAGAGCCCGGC





893
CCCCACCAAGAGCCCGGCG





894
CCCACCAAGAGCCCGGCGC





895
CCACCAAGAGCCCGGCGCC





896
CACCAAGAGCCCGGCGCCU





897
ACCAAGAGCCCGGCGCCUU





898
CCAAGAGCCCGGCGCCUUC





899
CAAGAGCCCGGCGCCUUCG





900
AAGAGCCCGGCGCCUUCGC





901
AGAGCCCGGCGCCUUCGCG





902
GAGCCCGGCGCCUUCGCGG





903
AGCCCGGCGCCUUCGCGGU





904
GCCCGGCGCCUUCGCGGUG





905
CCCGGCGCCUUCGCGGUGA





906
CCGGCGCCUUCGCGGUGAG





907
CGGCGCCUUCGCGGUGAGC





908
GGCGCCUUCGCGGUGAGCC





909
GCGCCUUCGCGGUGAGCCC





910
CGCCUUCGCGGUGAGCCCG





911
GCCUUCGCGGUGAGCCCGG





912
CCUUCGCGGUGAGCCCGGU





913
CUUCGCGGUGAGCCCGGUU





914
UUCGCGGUGAGCCCGGUUC





915
UCGCGGUGAGCCCGGUUCU





916
CGCGGUGAGCCCGGUUCUU





917
GCGGUGAGCCCGGUUCUUG





918
CGGUGAGCCCGGUUCUUGA





919
GGUGAGCCCGGUUCUUGAG





920
GUGAGCCCGGUUCUUGAGG





921
UGAGCCCGGUUCUUGAGGG





922
GAGCCCGGUUCUUGAGGGC





923
AGCCCGGUUCUUGAGGGCG





924
GCCCGGUUCUUGAGGGCGA





925
CCCGGUUCUUGAGGGCGAG





926
CCGGUUCUUGAGGGCGAGU





927
CGGUUCUUGAGGGCGAGUA





928
GGUUCUUGAGGGCGAGUAG





929
GUUCUUGAGGGCGAGUAGG





930
UUCUUGAGGGCGAGUAGGC





931
UCUUGAGGGCGAGUAGGCG





932
CUUGAGGGCGAGUAGGCGC





933
UUGAGGGCGAGUAGGCGCG





934
UGAGGGCGAGUAGGCGCGG





935
GAGGGCGAGUAGGCGCGGC





936
AGGGCGAGUAGGCGCGGCG





937
GGGCGAGUAGGCGCGGCGU





938
GGCGAGUAGGCGCGGCGUC





939
GCGAGUAGGCGCGGCGUCG





940
CGAGUAGGCGCGGCGUCGG





941
GAGUAGGCGCGGCGUCGGG





942
AGUAGGCGCGGCGUCGGGC





943
GUAGGCGCGGCGUCGGGCG





944
UAGGCGCGGCGUCGGGCGG





945
AGGCGCGGCGUCGGGCGGC





946
GGCGCGGCGUCGGGCGGCU





947
GCGCGGCGUCGGGCGGCUG





948
CGCGGCGUCGGGCGGCUGC





949
GCGGCGUCGGGCGGCUGCU





950
CGGCGUCGGGCGGCUGCUG





951
GGCGUCGGGCGGCUGCUGC





952
GCGUCGGGCGGCUGCUGCG





953
CGUCGGGCGGCUGCUGCGC





954
GUCGGGCGGCUGCUGCGCG





955
UCGGGCGGCUGCUGCGCGG





956
CGGGCGGCUGCUGCGCGGC





957
GGGCGGCUGCUGCGCGGCG





958
GGCGGCUGCUGCGCGGCGU





959
GCGGCUGCUGCGCGGCGUU





960
CGGCUGCUGCGCGGCGUUC





961
GGCUGCUGCGCGGCGUUCA





962
GCUGCUGCGCGGCGUUCAC





963
CUGCUGCGCGGCGUUCACU





964
UGCUGCGCGGCGUUCACUG





965
GCUGCGCGGCGUUCACUGU





966
CUGCGCGGCGUUCACUGUU





967
UGCGCGGCGUUCACUGUUG





968
GCGCGGCGUUCACUGUUGC





969
CGCGGCGUUCACUGUUGCC





970
GCGGCGUUCACUGUUGCCU





971
CGGCGUUCACUGUUGCCUU





972
GGCGUUCACUGUUGCCUUG





973
GCGUUCACUGUUGCCUUGU





974
CGUUCACUGUUGCCUUGUU





975
GUUCACUGUUGCCUUGUUC





976
UUCACUGUUGCCUUGUUCU





977
UCACUGUUGCCUUGUUCUG





978
CACUGUUGCCUUGUUCUGU





979
ACUGUUGCCUUGUUCUGUU





980
CUGUUGCCUUGUUCUGUUG





981
UGUUGCCUUGUUCUGUUGG





982
GUUGCCUUGUUCUGUUGGG





983
UUGCCUUGUUCUGUUGGGG





984
UGCCUUGUUCUGUUGGGGU





985
GCCUUGUUCUGUUGGGGUU





986
CCUUGUUCUGUUGGGGUUG





987
CUUGUUCUGUUGGGGUUGC





988
UUGUUCUGUUGGGGUUGCG





989
UGUUCUGUUGGGGUUGCGG





990
GUUCUGUUGGGGUUGCGGG





991
UUCUGUUGGGGUUGCGGGG





992
UCUGUUGGGGUUGCGGGGG





993
CUGUUGGGGUUGCGGGGGG





994
UGUUGGGGUUGCGGGGGGG





995
GUUGGGGUUGCGGGGGGGC





996
UUGGGGUUGCGGGGGGGCG





997
UGGGGUUGCGGGGGGGCGU





998
GGGGUUGCGGGGGGGCGUU





999
GGGUUGCGGGGGGGCGUUG





1000
GGUUGCGGGGGGGCGUUGG





1001
GUUGCGGGGGGGCGUUGGG





1002
UUGCGGGGGGGCGUUGGGU





1003
UGCGGGGGGGCGUUGGGUU





1004
GCGGGGGGGCGUUGGGUUU





1005
CGGGGGGGCGUUGGGUUUC





1006
GGGGGGGCGUUGGGUUUCU





1007
GGGGGGCGUUGGGUUUCUU





1008
GGGGGCGUUGGGUUUCUUC





1009
GGGGCGUUGGGUUUCUUCU





1010
GGGCGUUGGGUUUCUUCUU





1011
GGCGUUGGGUUUCUUCUUU





1012
GCGUUGGGUUUCUUCUUUC





1013
CGUUGGGUUUCUUCUUUCC





1014
GUUGGGUUUCUUCUUUCCG





1015
UUGGGUUUCUUCUUUCCGG





1016
UGGGUUUCUUCUUUCCGGG





1017
GGGUUUCUUCUUUCCGGGG





1018
GGUUUCUUCUUUCCGGGGC





1019
GUUUCUUCUUUCCGGGGCG





1020
UUUCUUCUUUCCGGGGCGG





1021
UUCUUCUUUCCGGGGCGGG





1022
UCUUCUUUCCGGGGCGGGG





1023
CUUCUUUCCGGGGCGGGGG





1024
UUCUUUCCGGGGCGGGGGG





1025
UCUUUCCGGGGCGGGGGGG





1026
CUUUCCGGGGCGGGGGGGG





1027
UUUCCGGGGCGGGGGGGGC





1028
UUCCGGGGCGGGGGGGGCA





1029
UCCGGGGCGGGGGGGGCAC





1030
CCGGGGCGGGGGGGGCACG





1031
CGGGGCGGGGGGGGCACGG





1032
GGGGCGGGGGGGGCACGGC





1033
GGGCGGGGGGGGCACGGCG





1034
GGCGGGGGGGGCACGGCGG





1035
GCGGGGGGGGCACGGCGGG





1036
CGGGGGGGGCACGGCGGGG





1037
GGGGGGGGCACGGCGGGGC





1038
GGGGGGGCACGGCGGGGCC





1039
GGGGGGCACGGCGGGGCCG





1040
GGGGGCACGGCGGGGCCGC





1041
GGGGCACGGCGGGGCCGCG





1042
GGGCACGGCGGGGCCGCGG





1043
GGCACGGCGGGGCCGCGGC





1044
GCACGGCGGGGCCGCGGCC





1045
CACGGCGGGGCCGCGGCCG





1046
ACGGCGGGGCCGCGGCCGG





1047
CGGCGGGGCCGCGGCCGGG





1048
GGCGGGGCCGCGGCCGGGC





1049
GCGGGGCCGCGGCCGGGCC





1050
CGGGGCCGCGGCCGGGCCG





1051
GGGGCCGCGGCCGGGCCGG





1052
GGGCCGCGGCCGGGCCGGC





1053
GGCCGCGGCCGGGCCGGCG





1054
GCCGCGGCCGGGCCGGCGG





1055
CCGCGGCCGGGCCGGCGGG





1056
CGCGGCCGGGCCGGCGGGG





1057
GCGGCCGGGCCGGCGGGGC





1058
CGGCCGGGCCGGCGGGGCG





1059
GGCCGGGCCGGCGGGGCGG





1060
GCCGGGCCGGCGGGGCGGG





1061
CCGGGCCGGCGGGGCGGGG





1062
CGGGCCGGCGGGGCGGGGC





1063
GGGCCGGCGGGGCGGGGCG





1064
GGCCGGCGGGGCGGGGCGG





1065
GCCGGCGGGGCGGGGCGGG





1066
CCGGCGGGGCGGGGCGGGG





1067
CGGCGGGGCGGGGCGGGGC





1068
GGCGGGGCGGGGCGGGGCG





1069
GCGGGGCGGGGCGGGGCGG





1070
CGGGGCGGGGCGGGGCGGG





1071
GGGGCGGGGCGGGGCGGGA





1072
GGGCGGGGCGGGGCGGGAC





1073
GGCGGGGCGGGGCGGGACG





1074
GCGGGGCGGGGCGGGACGG





1075
CGGGGCGGGGCGGGACGGG





1076
GGGGCGGGGCGGGACGGGG





1077
GGGCGGGGCGGGACGGGGC





1078
GGCGGGGCGGGACGGGGCG





1079
GCGGGGCGGGACGGGGCGG





1080
CGGGGCGGGACGGGGCGGG





1081
GGGGCGGGACGGGGCGGGG





1082
GGGCGGGACGGGGCGGGGC





1083
GGCGGGACGGGGCGGGGCG





1084
GCGGGACGGGGCGGGGCGG





1085
CGGGACGGGGCGGGGCGGA





1086
GGGACGGGGCGGGGCGGAG





1087
GGACGGGGCGGGGCGGAGC





1088
GACGGGGCGGGGCGGAGCC





1089
ACGGGGCGGGGCGGAGCCG





1090
CGGGGCGGGGCGGAGCCGC





1091
GGGGCGGGGCGGAGCCGCG





1092
GGGCGGGGCGGAGCCGCGC





1093
GGCGGGGCGGAGCCGCGCG





1094
GCGGGGCGGAGCCGCGCGG





1095
CGGGGCGGAGCCGCGCGGG





1096
GGGGCGGAGCCGCGCGGGG





1097
GGGCGGAGCCGCGCGGGGG





1098
GGCGGAGCCGCGCGGGGGC





1099
GCGGAGCCGCGCGGGGGCC





1100
CGGAGCCGCGCGGGGGCCG





1101
GGAGCCGCGCGGGGGCCGC





1102
GAGCCGCGCGGGGGCCGCA





1103
AGCCGCGCGGGGGCCGCAG





1104
GCCGCGCGGGGGCCGCAGU





1105
CCGCGCGGGGGCCGCAGUC





1106
CGCGCGGGGGCCGCAGUCC





1107
GCGCGGGGGCCGCAGUCCG





1108
CGCGGGGGCCGCAGUCCGG





1109
GCGGGGGCCGCAGUCCGGG





1110
CGGGGGCCGCAGUCCGGGC





1111
GGGGGCCGCAGUCCGGGCC





1112
GGGGCCGCAGUCCGGGCCG





1113
GGGCCGCAGUCCGGGCCGG





1114
GGCCGCAGUCCGGGCCGGG





1115
GCCGCAGUCCGGGCCGGGG





1116
CCGCAGUCCGGGCCGGGGC





1117
CGCAGUCCGGGCCGGGGCC





1118
GCAGUCCGGGCCGGGGCCG





1119
CAGUCCGGGCCGGGGCCGC





1120
AGUCCGGGCCGGGGCCGCC





1121
GUCCGGGCCGGGGCCGCCG





1122
UCCGGGCCGGGGCCGCCGU





1123
CCGGGCCGGGGCCGCCGUC





1124
CGGGCCGGGGCCGCCGUCG





1125
GGGCCGGGGCCGCCGUCGG





1126
GGCCGGGGCCGCCGUCGGG





1127
GCCGGGGCCGCCGUCGGGU





1128
CCGGGGCCGCCGUCGGGUC





1129
CGGGGCCGCCGUCGGGUCU





1130
GGGGCCGCCGUCGGGUCUC





1131
GGGCCGCCGUCGGGUCUCG





1132
GGCCGCCGUCGGGUCUCGG





1133
GCCGCCGUCGGGUCUCGGC





1134
CCGCCGUCGGGUCUCGGCC





1135
CGCCGUCGGGUCUCGGCCC





1136
GCCGUCGGGUCUCGGCCCG





1137
CCGUCGGGUCUCGGCCCGC





1138
CGUCGGGUCUCGGCCCGCU





1139
GUCGGGUCUCGGCCCGCUC





1140
UCGGGUCUCGGCCCGCUCC





1141
CGGGUCUCGGCCCGCUCCC





1142
GGGUCUCGGCCCGCUCCCG





1143
GGUCUCGGCCCGCUCCCGU





1144
GUCUCGGCCCGCUCCCGUC





1145
UCUCGGCCCGCUCCCGUCG





1146
CUCGGCCCGCUCCCGUCGG





1147
UCGGCCCGCUCCCGUCGGG





1148
CGGCCCGCUCCCGUCGGGG





1149
GGCCCGCUCCCGUCGGGGC





1150
GCCCGCUCCCGUCGGGGCG





1151
CCCGCUCCCGUCGGGGCGG





1152
CCGCUCCCGUCGGGGCGGA





1153
CGCUCCCGUCGGGGCGGAG





1154
GCUCCCGUCGGGGCGGAGC





1155
CUCCCGUCGGGGCGGAGCG





1156
UCCCGUCGGGGCGGAGCGU





1157
CCCGUCGGGGCGGAGCGUC





1158
CCGUCGGGGCGGAGCGUCC





1159
CGUCGGGGCGGAGCGUCCG





1160
GUCGGGGCGGAGCGUCCGA





1161
UCGGGGCGGAGCGUCCGAC





1162
CGGGGCGGAGCGUCCGACG





1163
GGGGCGGAGCGUCCGACGA





1164
GGGCGGAGCGUCCGACGAU





1165
GGCGGAGCGUCCGACGAUC





1166
GCGGAGCGUCCGACGAUCG





1167
CGGAGCGUCCGACGAUCGG





1168
GGAGCGUCCGACGAUCGGC





1169
GAGCGUCCGACGAUCGGCC





1170
AGCGUCCGACGAUCGGCCU





1171
GCGUCCGACGAUCGGCCUC





1172
CGUCCGACGAUCGGCCUCC





1173
GUCCGACGAUCGGCCUCCA





1174
UCCGACGAUCGGCCUCCAC





1175
CCGACGAUCGGCCUCCACG





1176
CGACGAUCGGCCUCCACGA





1177
GACGAUCGGCCUCCACGAA





1178
ACGAUCGGCCUCCACGAAA





1179
CGAUCGGCCUCCACGAAAC





1180
GAUCGGCCUCCACGAAACG





1181
AUCGGCCUCCACGAAACGC





1182
UCGGCCUCCACGAAACGCG





1183
CGGCCUCCACGAAACGCGG





1184
GGCCUCCACGAAACGCGGU





1185
GCCUCCACGAAACGCGGUG





1186
CCUCCACGAAACGCGGUGC





1187
CUCCACGAAACGCGGUGCC





1188
UCCACGAAACGCGGUGCCG





1189
CCACGAAACGCGGUGCCGU





1190
CACGAAACGCGGUGCCGUG





1191
ACGAAACGCGGUGCCGUGA





1192
CGAAACGCGGUGCCGUGAU





1193
GAAACGCGGUGCCGUGAUG





1194
AAACGCGGUGCCGUGAUGU





1195
AACGCGGUGCCGUGAUGUG





1196
ACGCGGUGCCGUGAUGUGU





1197
CGCGGUGCCGUGAUGUGUU





1198
GCGGUGCCGUGAUGUGUUU





1199
CGGUGCCGUGAUGUGUUUG





1200
GGUGCCGUGAUGUGUUUGU





1201
GUGCCGUGAUGUGUUUGUA





1202
UGCCGUGAUGUGUUUGUAG





1203
GCCGUGAUGUGUUUGUAGU





1204
CCGUGAUGUGUUUGUAGUG





1205
CGUGAUGUGUUUGUAGUGG





1206
GUGAUGUGUUUGUAGUGGU





1207
UGAUGUGUUUGUAGUGGUU





1208
GAUGUGUUUGUAGUGGUUC





1209
AUGUGUUUGUAGUGGUUCC





1210
UGUGUUUGUAGUGGUUCCU





1211
GUGUUUGUAGUGGUUCCUC





1212
UGUUUGUAGUGGUUCCUCG





1213
GUUUGUAGUGGUUCCUCGU





1214
UUUGUAGUGGUUCCUCGUA





1215
UUGUAGUGGUUCCUCGUAG





1216
UGUAGUGGUUCCUCGUAGG





1217
GUAGUGGUUCCUCGUAGGC





1218
UAGUGGUUCCUCGUAGGCU





1219
AGUGGUUCCUCGUAGGCUC





1220
GUGGUUCCUCGUAGGCUCC





1221
UGGUUCCUCGUAGGCUCCA





1222
GGUUCCUCGUAGGCUCCAG





1223
GUUCCUCGUAGGCUCCAGA





1224
UUCCUCGUAGGCUCCAGAC





1225
UCCUCGUAGGCUCCAGACG





1226
CCUCGUAGGCUCCAGACGU





1227
CUCGUAGGCUCCAGACGUU





1228
UCGUAGGCUCCAGACGUUU





1229
CGUAGGCUCCAGACGUUUU





1230
GUAGGCUCCAGACGUUUUC





1231
UAGGCUCCAGACGUUUUCU





1232
AGGCUCCAGACGUUUUCUC





1233
GGCUCCAGACGUUUUCUCC





1234
GCUCCAGACGUUUUCUCCU





1235
CUCCAGACGUUUUCUCCUC





1236
UCCAGACGUUUUCUCCUCG





1237
CCAGACGUUUUCUCCUCGU





1238
CAGACGUUUUCUCCUCGUA





1239
AGACGUUUUCUCCUCGUAU





1240
GACGUUUUCUCCUCGUAUC





1241
ACGUUUUCUCCUCGUAUCG





1242
CGUUUUCUCCUCGUAUCGC





1243
GUUUUCUCCUCGUAUCGCC





1244
UUUUCUCCUCGUAUCGCCA





1245
UUUCUCCUCGUAUCGCCAA





1246
UUCUCCUCGUAUCGCCAAA





1247
UCUCCUCGUAUCGCCAAAU





1248
CUCCUCGUAUCGCCAAAUU





1249
UCCUCGUAUCGCCAAAUUA





1250
CCUCGUAUCGCCAAAUUAA





1251
CUCGUAUCGCCAAAUUAAC





1252
UCGUAUCGCCAAAUUAACG





1253
CGUAUCGCCAAAUUAACGC





1254
GUAUCGCCAAAUUAACGCG





1255
UAUCGCCAAAUUAACGCGU





1256
AUCGCCAAAUUAACGCGUU





1257
UCGCCAAAUUAACGCGUUU





1258
CGCCAAAUUAACGCGUUUU





1259
GCCAAAUUAACGCGUUUUG





1260
CCAAAUUAACGCGUUUUGC





1261
CAAAUUAACGCGUUUUGCA





1262
AAAUUAACGCGUUUUGCAU





1263
AAUUAACGCGUUUUGCAUA





1264
AUUAACGCGUUUUGCAUAU





1265
UUAACGCGUUUUGCAUAUU





1266
UAACGCGUUUUGCAUAUUA





1267
AACGCGUUUUGCAUAUUAC





1268
ACGCGUUUUGCAUAUUACA





1269
CGCGUUUUGCAUAUUACAG





1270
GCGUUUUGCAUAUUACAGU





1271
CGUUUUGCAUAUUACAGUU





1272
GUUUUGCAUAUUACAGUUG





1273
UUUUGCAUAUUACAGUUGA





1274
UUUGCAUAUUACAGUUGAG





1275
UUGCAUAUUACAGUUGAGU





1276
UGCAUAUUACAGUUGAGUG





1277
GCAUAUUACAGUUGAGUGC





1278
CAUAUUACAGUUGAGUGCC





1279
AUAUUACAGUUGAGUGCCU





1280
UAUUACAGUUGAGUGCCUC





1281
AUUACAGUUGAGUGCCUCG





1282
UUACAGUUGAGUGCCUCGA





1283
UACAGUUGAGUGCCUCGAC





1284
ACAGUUGAGUGCCUCGACU





1285
CAGUUGAGUGCCUCGACUU





1286
AGUUGAGUGCCUCGACUUA





1287
GUUGAGUGCCUCGACUUAG





1288
UUGAGUGCCUCGACUUAGA





1289
UGAGUGCCUCGACUUAGAU





1290
GAGUGCCUCGACUUAGAUU





1291
AGUGCCUCGACUUAGAUUG





1292
GUGCCUCGACUUAGAUUGC





1293
UGCCUCGACUUAGAUUGCA





1294
GCCUCGACUUAGAUUGCAA





1295
CCUCGACUUAGAUUGCAAU





1296
CUCGACUUAGAUUGCAAUA





1297
UCGACUUAGAUUGCAAUAU





1298
CGACUUAGAUUGCAAUAUA





1299
GACUUAGAUUGCAAUAUAA





1300
ACUUAGAUUGCAAUAUAAG





1301
CUUAGAUUGCAAUAUAAGC





1302
UUAGAUUGCAAUAUAAGCG





1303
UAGAUUGCAAUAUAAGCGG





1304
AGAUUGCAAUAUAAGCGGC





1305
GAUUGCAAUAUAAGCGGCC





1306
AUUGCAAUAUAAGCGGCCA





1307
UUGCAAUAUAAGCGGCCAG





1308
UGCAAUAUAAGCGGCCAGC





1309
GCAAUAUAAGCGGCCAGCA





1310
CAAUAUAAGCGGCCAGCAA





1311
AAUAUAAGCGGCCAGCAAA





1312
AUAUAAGCGGCCAGCAAAC





1313
UAUAAGCGGCCAGCAAACA





1314
AUAAGCGGCCAGCAAACAA





1315
UAAGCGGCCAGCAAACAAG





1316
AAGCGGCCAGCAAACAAGU





1317
AGCGGCCAGCAAACAAGUC





1318
GCGGCCAGCAAACAAGUCU





1319
CGGCCAGCAAACAAGUCUC





1320
GGCCAGCAAACAAGUCUCA





1321
GCCAGCAAACAAGUCUCAA





1322
CCAGCAAACAAGUCUCAAA





1323
CAGCAAACAAGUCUCAAAA





1324
AGCAAACAAGUCUCAAAAA





1325
GCAAACAAGUCUCAAAAAA





1326
CAAACAAGUCUCAAAAAAA





1327
AAACAAGUCUCAAAAAAAA





1328
AACAAGUCUCAAAAAAAAG





1329
ACAAGUCUCAAAAAAAAGU





1330
CAAGUCUCAAAAAAAAGUU





1331
AAGUCUCAAAAAAAAGUUA





1332
AGUCUCAAAAAAAAGUUAC





1333
GUCUCAAAAAAAAGUUACG





1334
UCUCAAAAAAAAGUUACGU





1335
CUCAAAAAAAAGUUACGUG





1336
UCAAAAAAAAGUUACGUGC





1337
CAAAAAAAAGUUACGUGCG





1338
AAAAAAAAGUUACGUGCGU





1339
AAAAAAAGUUACGUGCGUU





1340
AAAAAAGUUACGUGCGUUU





1341
AAAAAGUUACGUGCGUUUC





1342
AAAAGUUACGUGCGUUUCU





1343
AAAGUUACGUGCGUUUCUG





1344
AAGUUACGUGCGUUUCUGC





1345
AGUUACGUGCGUUUCUGCG





1346
GUUACGUGCGUUUCUGCGA





1347
UUACGUGCGUUUCUGCGAG





1348
UACGUGCGUUUCUGCGAGU





1349
ACGUGCGUUUCUGCGAGUG





1350
CGUGCGUUUCUGCGAGUGU





1351
GUGCGUUUCUGCGAGUGUU





1352
UGCGUUUCUGCGAGUGUUA





1353
GCGUUUCUGCGAGUGUUAU





1354
CGUUUCUGCGAGUGUUAUU





1355
GUUUCUGCGAGUGUUAUUU





1356
UUUCUGCGAGUGUUAUUUU





1357
UUCUGCGAGUGUUAUUUUG





1358
UCUGCGAGUGUUAUUUUGU





1359
CUGCGAGUGUUAUUUUGUU





1360
UGCGAGUGUUAUUUUGUUA





1361
GCGAGUGUUAUUUUGUUAA





1362
CGAGUGUUAUUUUGUUAAG





1363
GAGUGUUAUUUUGUUAAGA





1364
AGUGUUAUUUUGUUAAGAA





1365
GUGUUAUUUUGUUAAGAAC





1366
UGUUAUUUUGUUAAGAACG





1367
GUUAUUUUGUUAAGAACGG





1368
UUAUUUUGUUAAGAACGGC





1369
UAUUUUGUUAAGAACGGCU





1370
AUUUUGUUAAGAACGGCUC





1371
UUUUGUUAAGAACGGCUCA





1372
UUUGUUAAGAACGGCUCAC





1373
UUGUUAAGAACGGCUCACA





1374
UGUUAAGAACGGCUCACAG





1375
GUUAAGAACGGCUCACAGU





1376
UUAAGAACGGCUCACAGUG





1377
UAAGAACGGCUCACAGUGU





1378
AAGAACGGCUCACAGUGUC





1379
AGAACGGCUCACAGUGUCC





1380
GAACGGCUCACAGUGUCCU





1381
AACGGCUCACAGUGUCCUC





1382
ACGGCUCACAGUGUCCUCU





1383
CGGCUCACAGUGUCCUCUU





1384
GGCUCACAGUGUCCUCUUC





1385
GCUCACAGUGUCCUCUUCC





1386
CUCACAGUGUCCUCUUCCU





1387
UCACAGUGUCCUCUUCCUG





1388
CACAGUGUCCUCUUCCUGU





1389
ACAGUGUCCUCUUCCUGUG





1390
CAGUGUCCUCUUCCUGUGU





1391
AGUGUCCUCUUCCUGUGUU





1392
GUGUCCUCUUCCUGUGUUA





1393
UGUCCUCUUCCUGUGUUAC





1394
GUCCUCUUCCUGUGUUACA





1395
UCCUCUUCCUGUGUUACAG





1396
CCUCUUCCUGUGUUACAGA





1397
CUCUUCCUGUGUUACAGAA





1398
UCUUCCUGUGUUACAGAAG





1399
CUUCCUGUGUUACAGAAGC





1400
UUCCUGUGUUACAGAAGCC





1401
UCCUGUGUUACAGAAGCCA





1402
CCUGUGUUACAGAAGCCAA





1403
CUGUGUUACAGAAGCCAAC





1404
UGUGUUACAGAAGCCAACC





1405
GUGUUACAGAAGCCAACCU





1406
UGUUACAGAAGCCAACCUG





1407
GUUACAGAAGCCAACCUGA





1408
UUACAGAAGCCAACCUGAA





1409
UACAGAAGCCAACCUGAAA





1410
ACAGAAGCCAACCUGAAAU





1411
CAGAAGCCAACCUGAAAUG





1412
AGAAGCCAACCUGAAAUGA





1413
GAAGCCAACCUGAAAUGAA





1414
AAGCCAACCUGAAAUGAAA





1415
AGCCAACCUGAAAUGAAAC





1416
GCCAACCUGAAAUGAAACU





1417
CCAACCUGAAAUGAAACUA





1418
CAACCUGAAAUGAAACUAG





1419
AACCUGAAAUGAAACUAGU





1420
ACCUGAAAUGAAACUAGUC





1421
CCUGAAAUGAAACUAGUCU





1422
CUGAAAUGAAACUAGUCUG





1423
UGAAAUGAAACUAGUCUGG





1424
GAAAUGAAACUAGUCUGGA





1425
AAAUGAAACUAGUCUGGAA





1426
AAUGAAACUAGUCUGGAAA





1427
AUGAAACUAGUCUGGAAAA





1428
UGAAACUAGUCUGGAAAAA





1429
GAAACUAGUCUGGAAAAAU





1430
AAACUAGUCUGGAAAAAUU





1431
AACUAGUCUGGAAAAAUUC





1432
ACUAGUCUGGAAAAAUUCA





1433
CUAGUCUGGAAAAAUUCAU





1434
UAGUCUGGAAAAAUUCAUU





1435
AGUCUGGAAAAAUUCAUUG





1436
GUCUGGAAAAAUUCAUUGU





1437
UCUGGAAAAAUUCAUUGUU





1438
CUGGAAAAAUUCAUUGUUC





1439
UGGAAAAAUUCAUUGUUCU





1440
GGAAAAAUUCAUUGUUCUC





1441
GAAAAAUUCAUUGUUCUCU





1442
AAAAAUUCAUUGUUCUCUG





1443
AAAAUUCAUUGUUCUCUGU





1444
AAAUUCAUUGUUCUCUGUA





1445
AAUUCAUUGUUCUCUGUAG





1446
AUUCAUUGUUCUCUGUAGU





1447
UUCAUUGUUCUCUGUAGUU





1448
UCAUUGUUCUCUGUAGUUG





1449
CAUUGUUCUCUGUAGUUGC





1450
AUUGUUCUCUGUAGUUGCA





1451
UUGUUCUCUGUAGUUGCAG





1452
UGUUCUCUGUAGUUGCAGC





1453
GUUCUCUGUAGUUGCAGCU





1454
UUCUCUGUAGUUGCAGCUG





1455
UCUCUGUAGUUGCAGCUGU





1456
CUCUGUAGUUGCAGCUGUA





1457
UCUGUAGUUGCAGCUGUAC





1458
CUGUAGUUGCAGCUGUACC





1459
UGUAGUUGCAGCUGUACCU





1460
GUAGUUGCAGCUGUACCUG





1461
UAGUUGCAGCUGUACCUGA





1462
AGUUGCAGCUGUACCUGAA





1463
GUUGCAGCUGUACCUGAAA





1464
UUGCAGCUGUACCUGAAAU





1465
UGCAGCUGUACCUGAAAUA





1466
GCAGCUGUACCUGAAAUAA





1467
CAGCUGUACCUGAAAUAAA





1468
AGCUGUACCUGAAAUAAAA





1469
GCUGUACCUGAAAUAAAAA





1470
CUGUACCUGAAAUAAAAAU





1471
UGUACCUGAAAUAAAAAUG





1472
GUACCUGAAAUAAAAAUGU





1473
UACCUGAAAUAAAAAUGUU





1474
ACCUGAAAUAAAAAUGUUA





1475
CCUGAAAUAAAAAUGUUAU





1476
CUGAAAUAAAAAUGUUAUU





1477
UGAAAUAAAAAUGUUAUUG





1478
GAAAUAAAAAUGUUAUUGA





1479
AAAUAAAAAUGUUAUUGAU





1480
AAUAAAAAUGUUAUUGAUG





1481
AUAAAAAUGUUAUUGAUGA





1482
UAAAAAUGUUAUUGAUGAC





1483
AAAAAUGUUAUUGAUGACU





1484
AAAAUGUUAUUGAUGACUG





1485
AAAUGUUAUUGAUGACUGA





1486
AAUGUUAUUGAUGACUGAA





1487
AUGUUAUUGAUGACUGAAA





1488
UGUUAUUGAUGACUGAAAA





1489
GUUAUUGAUGACUGAAAAA





1490
UUAUUGAUGACUGAAAAAA





1491
UAUUGAUGACUGAAAAAAA





1492
AUUGAUGACUGAAAAAAAA





1493
UUGAUGACUGAAAAAAAA





1494
UGAUGACUGAAAAAAAAA





1495
GAUGACUGAAAAAAAAAA





1496
AUGACUGAAAAAAAAAAA





1497
UGACUGAAAAAAAAAAAA





1498
GACUGAAAAAAAAAAAAA





1499
ACUGAAAAAAAAAAAAAA





1500
CUGAAAAAAAAAAAAAAA





1501
UGAAAAAAAAAAAAAAAA





1502
AAAACAGUGGCAGAUGAAA





1503
AAACAGUGGCAGAUGAAAG





1504
AACAGUGGCAGAUGAAAGG





1505
ACAGUGGCAGAUGAAAGGG





1506
CAGUGGCAGAUGAAAGGGA





1507
AGUGGCAGAUGAAAGGGAU





1508
GUGGCAGAUGAAAGGGAUG





1509
UGGCAGAUGAAAGGGAUGG





1510
GGCAGAUGAAAGGGAUGGA





1511
GCAGAUGAAAGGGAUGGAG





1512
CAGAUGAAAGGGAUGGAGA





1513
AGAUGAAAGGGAUGGAGAA





1514
GAUGAAAGGGAUGGAGAAC





1515
AUGAAAGGGAUGGAGAACC





1516
UGAAAGGGAUGGAGAACCG





1517
GAAAGGGAUGGAGAACCGC





1518
AAAGGGAUGGAGAACCGCC





1519
AAGGGAUGGAGAACCGCCA





1520
AGGGAUGGAGAACCGCCAU





1521
GGGAUGGAGAACCGCCAUG





1522
GGAUGGAGAACCGCCAUGC





1523
GAUGGAGAACCGCCAUGCC





1524
AUGCCCGGUGACUCCCCGG





1525
UGCCCGGUGACUCCCCGGC





1526
GCCCGGUGACUCCCCGGCC





1527
CCCGGUGACUCCCCGGCCG





1528
CCGGUGACUCCCCGGCCGU





1529
CGGUGACUCCCCGGCCGUC





1530
GGUGACUCCCCGGCCGUCA





1531
GUGACUCCCCGGCCGUCAG





1532
UGACUCCCCGGCCGUCAGC





1533
GACUCCCCGGCCGUCAGCA





1534
ACUCCCCGGCCGUCAGCAA





1535
CUCCCCGGCCGUCAGCAAG





1536
UCCCCGGCCGUCAGCAAGC





1537
CCCCGGCCGUCAGCAAGCC





1538
CCCGGCCGUCAGCAAGCCC





1539
CCGGCCGUCAGCAAGCCCC





1540
CGGCCGUCAGCAAGCCCCC





1541
GGCCGUCAGCAAGCCCCCG





1542
GCCGUCAGCAAGCCCCCGG





1543
CCGUCAGCAAGCCCCCGGA





1544
CGUCAGCAAGCCCCCGGAC





1545
GUCAGCAAGCCCCCGGACG





1546
UCAGCAAGCCCCCGGACGG





1547
CAGCAAGCCCCCGGACGGC





1548
AGCAAGCCCCCGGACGGCG





1549
GCAAGCCCCCGGACGGCGC





1550
CAAGCCCCCGGACGGCGCC





1551
AAGCCCCCGGACGGCGCCG





1552
AGCCCCCGGACGGCGCCGG





1553
GCCCCCGGACGGCGCCGGG





1554
CCCCCGGACGGCGCCGGGC





1555
CCCCGGACGGCGCCGGGCC





1556
CCCGGACGGCGCCGGGCCG





1557
CCGGACGGCGCCGGGCCGG





1558
CGGACGGCGCCGGGCCGGG





1559
GGACGGCGCCGGGCCGGGG





1560
GACGGCGCCGGGCCGGGGG





1561
ACGGCGCCGGGCCGGGGGA





1562
CGGCGCCGGGCCGGGGGAC





1563
GGCGCCGGGCCGGGGGACA





1564
GCGCCGGGCCGGGGGACAA





1565
CGCCGGGCCGGGGGACAAG





1566
GCCGGGCCGGGGGACAAGG





1567
CCGGGCCGGGGGACAAGGC





1568
CGGGCCGGGGGACAAGGCG





1569
GGGCCGGGGGACAAGGCGG





1570
GGCCGGGGGACAAGGCGGG





1571
GCCGGGGGACAAGGCGGGC





1572
CCGGGGGACAAGGCGGGCG





1573
CGGGGGACAAGGCGGGCGG





1574
GGGGGACAAGGCGGGCGGC





1575
GGGGACAAGGCGGGCGGCC





1576
GGGACAAGGCGGGCGGCCU





1577
GGACAAGGCGGGCGGCCUC





1578
GACAAGGCGGGCGGCCUCG





1579
ACAAGGCGGGCGGCCUCGG





1580
CAAGGCGGGCGGCCUCGGC





1581
AAGGCGGGCGGCCUCGGCA





1582
AGGCGGGCGGCCUCGGCAA





1583
GGCGGGCGGCCUCGGCAAG





1584
GCGGGCGGCCUCGGCAAGG





1585
CGGGCGGCCUCGGCAAGGC





1586
GGGCGGCCUCGGCAAGGCG





1587
GGCGGCCUCGGCAAGGCGG





1588
GCGGCCUCGGCAAGGCGGC





1589
CGGCCUCGGCAAGGCGGCG





1590
GGCCUCGGCAAGGCGGCGG





1591
GCCUCGGCAAGGCGGCGGC





1592
CCUCGGCAAGGCGGCGGCC





1593
CUCGGCAAGGCGGCGGCCC





1594
UCGGCAAGGCGGCGGCCCA





1595
CGGCAAGGCGGCGGCCCAA





1596
GGCAAGGCGGCGGCCCAAA





1597
GCAAGGCGGCGGCCCAAAU





1598
CAAGGCGGCGGCCCAAAUG





1599
AAGGCGGCGGCCCAAAUGG





1600
AGGCGGCGGCCCAAAUGGC





1601
GGCGGCGGCCCAAAUGGCG





1602
GCGGCGGCCCAAAUGGCGG





1603
CGGCGGCCCAAAUGGCGGC





1604
GGCGGCCCAAAUGGCGGCC





1605
GCGGCCCAAAUGGCGGCCG





1606
CGGCCCAAAUGGCGGCCGC





1607
GGCCCAAAUGGCGGCCGCC





1608
GCCCAAAUGGCGGCCGCCC





1609
CCCAAAUGGCGGCCGCCCC





1610
CCAAAUGGCGGCCGCCCCG





1611
CAAAUGGCGGCCGCCCCGG





1612
AAAUGGCGGCCGCCCCGGC





1613
AAUGGCGGCCGCCCCGGCG





1614
AUGGCGGCCGCCCCGGCGG





1615
UGGCGGCCGCCCCGGCGGC





1616
GGCGGCCGCCCCGGCGGCG





1617
GCGGCCGCCCCGGCGGCGG





1618
CGGCCGCCCCGGCGGCGGG





1619
GGCCGCCCCGGCGGCGGGC





1620
GCCGCCCCGGCGGCGGGCA





1621
CCGCCCCGGCGGCGGGCAA





1622
CGCCCCGGCGGCGGGCAAG





1623
GCCCCGGCGGCGGGCAAGA





1624
CCCCGGCGGCGGGCAAGAA





1625
CCCGGCGGCGGGCAAGAAG





1626
GGCAGCGGGUGAUGGCCGC





1627
GCAGCGGGUGAUGGCCGCG





1628
CAGCGGGUGAUGGCCGCGC





1629
AGCGGGUGAUGGCCGCGCA





1630
GCGGGUGAUGGCCGCGCAG





1631
CGGGUGAUGGCCGCGCAGG





1632
GGGUGAUGGCCGCGCAGGU





1633
GGUGAUGGCCGCGCAGGUU





1634
GUGAUGGCCGCGCAGGUUG





1635
UGAUGGCCGCGCAGGUUGC





1636
GAUGGCCGCGCAGGUUGCA





1637
AUGGCCGCGCAGGUUGCAC





1638
UGGCCGCGCAGGUUGCACU





1639
GGCCGCGCAGGUUGCACUG





1640
GCCGCGCAGGUUGCACUGA





1641
CCGCGCAGGUUGCACUGAG





1642
CGCGCAGGUUGCACUGAGG





1643
GCGCAGGUUGCACUGAGGA





1644
CGCAGGUUGCACUGAGGAG
















TABLE 2







DsRNA molecules targeting mRNA encoding chicken


ASW (WPKCI).








SEQ ID NO
Sequence 5′-3′





1645
CGUCUGCGGGUGCCUUGCG





1646
GUCUGCGGGUGCCUUGCGA





1647
UCUGCGGGUGCCUUGCGAU





1648
CUGCGGGUGCCUUGCGAUA





1649
UGCGGGUGCCUUGCGAUAC





1650
GCGGGUGCCUUGCGAUACG





1651
CGGGUGCCUUGCGAUACGU





1652
GGGUGCCUUGCGAUACGUC





1653
GGUGCCUUGCGAUACGUCG





1654
GUGCCUUGCGAUACGUCGG





1655
UGCCUUGCGAUACGUCGGC





1656
GCCUUGCGAUACGUCGGCA





1657
CCUUGCGAUACGUCGGCAU





1658
CUUGCGAUACGUCGGCAUG





1659
UUGCGAUACGUCGGCAUGG





1660
UGCGAUACGUCGGCAUGGC





1661
GCGAUACGUCGGCAUGGCU





1662
CGAUACGUCGGCAUGGCUG





1663
GAUACGUCGGCAUGGCUGA





1664
AUACGUCGGCAUGGCUGAC





1665
UACGUCGGCAUGGCUGACG





1666
ACGUCGGCAUGGCUGACGA





1667
CGUCGGCAUGGCUGACGAG





1668
GUCGGCAUGGCUGACGAGA





1669
UCGGCAUGGCUGACGAGAU





1670
CGGCAUGGCUGACGAGAUC





1671
GGCAUGGCUGACGAGAUCC





1672
GCAUGGCUGACGAGAUCCG





1673
CAUGGCUGACGAGAUCCGC





1674
AUGGCUGACGAGAUCCGCA





1675
UGGCUGACGAGAUCCGCAA





1676
GGCUGACGAGAUCCGCAAG





1677
GCUGACGAGAUCCGCAAGG





1678
CUGACGAGAUCCGCAAGGC





1679
UGACGAGAUCCGCAAGGCG





1680
GACGAGAUCCGCAAGGCGC





1681
ACGAGAUCCGCAAGGCGCA





1682
CGAGAUCCGCAAGGCGCAG





1683
GAGAUCCGCAAGGCGCAGG





1684
AGAUCCGCAAGGCGCAGGC





1685
GAUCCGCAAGGCGCAGGCC





1686
AUCCGCAAGGCGCAGGCCG





1687
UCCGCAAGGCGCAGGCCGC





1688
CCGCAAGGCGCAGGCCGCG





1689
CGCAAGGCGCAGGCCGCGC





1690
GCAAGGCGCAGGCCGCGCG





1691
CAAGGCGCAGGCCGCGCGC





1692
AAGGCGCAGGCCGCGCGCC





1693
AGGCGCAGGCCGCGCGCCC





1694
GGCGCAGGCCGCGCGCCCU





1695
GCGCAGGCCGCGCGCCCUG





1696
CGCAGGCCGCGCGCCCUGG





1697
GCAGGCCGCGCGCCCUGGU





1698
CAGGCCGCGCGCCCUGGUG





1699
AGGCCGCGCGCCCUGGUGG





1700
GGCCGCGCGCCCUGGUGGG





1701
GCCGCGCGCCCUGGUGGGG





1702
CCGCGCGCCCUGGUGGGGA





1703
CGCGCGCCCUGGUGGGGAC





1704
GCGCGCCCUGGUGGGGACA





1705
CGCGCCCUGGUGGGGACAC





1706
GCGCCCUGGUGGGGACACC





1707
CGCCCUGGUGGGGACACCA





1708
GCCCUGGUGGGGACACCAU





1709
CCCUGGUGGGGACACCAUC





1710
CCUGGUGGGGACACCAUCU





1711
CUGGUGGGGACACCAUCUU





1712
UGGUGGGGACACCAUCUUC





1713
GGUGGGGACACCAUCUUCG





1714
GUGGGGACACCAUCUUCGG





1715
UGGGGACACCAUCUUCGGG





1716
GGGGACACCAUCUUCGGGA





1717
GGGACACCAUCUUCGGGAA





1718
GGACACCAUCUUCGGGAAG





1719
GACACCAUCUUCGGGAAGA





1720
ACACCAUCUUCGGGAAGAU





1721
CACCAUCUUCGGGAAGAUU





1722
ACCAUCUUCGGGAAGAUUA





1723
CCAUCUUCGGGAAGAUUAU





1724
CAUCUUCGGGAAGAUUAUC





1725
AUCUUCGGGAAGAUUAUCC





1726
UCUUCGGGAAGAUUAUCCG





1727
CUUCGGGAAGAUUAUCCGC





1728
UUCGGGAAGAUUAUCCGCA





1729
UCGGGAAGAUUAUCCGCAA





1730
CGGGAAGAUUAUCCGCAAG





1731
GGGAAGAUUAUCCGCAAGG





1732
GGAAGAUUAUCCGCAAGGA





1733
GAAGAUUAUCCGCAAGGAG





1734
AAGAUUAUCCGCAAGGAGA





1735
AGAUUAUCCGCAAGGAGAU





1736
GAUUAUCCGCAAGGAGAUU





1737
AUUAUCCGCAAGGAGAUUC





1738
UUAUCCGCAAGGAGAUUCC





1739
UAUCCGCAAGGAGAUUCCC





1740
AUCCGCAAGGAGAUUCCCG





1741
UCCGCAAGGAGAUUCCCGC





1742
CCGCAAGGAGAUUCCCGCC





1743
CGCAAGGAGAUUCCCGCCA





1744
GCAAGGAGAUUCCCGCCAA





1745
CAAGGAGAUUCCCGCCAAC





1746
AAGGAGAUUCCCGCCAACA





1747
AGGAGAUUCCCGCCAACAU





1748
GGAGAUUCCCGCCAACAUA





1749
GAGAUUCCCGCCAACAUAA





1750
AGAUUCCCGCCAACAUAAU





1751
GAUUCCCGCCAACAUAAUC





1752
AUUCCCGCCAACAUAAUCU





1753
UUCCCGCCAACAUAAUCUA





1754
UCCCGCCAACAUAAUCUAC





1755
CCCGCCAACAUAAUCUACG





1756
CCGCCAACAUAAUCUACGA





1757
CGCCAACAUAAUCUACGAG





1758
GCCAACAUAAUCUACGAGG





1759
CCAACAUAAUCUACGAGGA





1760
CAACAUAAUCUACGAGGAC





1761
AACAUAAUCUACGAGGACG





1762
ACAUAAUCUACGAGGACGA





1763
CAUAAUCUACGAGGACGAG





1764
AUAAUCUACGAGGACGAGC





1765
UAAUCUACGAGGACGAGCA





1766
AAUCUACGAGGACGAGCAG





1767
AUCUACGAGGACGAGCAGU





1768
UCUACGAGGACGAGCAGUG





1769
CUACGAGGACGAGCAGUGC





1770
UACGAGGACGAGCAGUGCC





1771
ACGAGGACGAGCAGUGCCU





1772
CGAGGACGAGCAGUGCCUU





1773
GAGGACGAGCAGUGCCUUG





1774
AGGACGAGCAGUGCCUUGC





1775
GGACGAGCAGUGCCUUGCG





1776
GACGAGCAGUGCCUUGCGU





1777
ACGAGCAGUGCCUUGCGUU





1778
CGAGCAGUGCCUUGCGUUC





1779
GAGCAGUGCCUUGCGUUCC





1780
AGCAGUGCCUUGCGUUCCA





1781
GCAGUGCCUUGCGUUCCAU





1782
CAGUGCCUUGCGUUCCAUG





1783
AGUGCCUUGCGUUCCAUGA





1784
GUGCCUUGCGUUCCAUGAU





1785
UGCCUUGCGUUCCAUGAUA





1786
GCCUUGCGUUCCAUGAUAU





1787
CCUUGCGUUCCAUGAUAUC





1788
CUUGCGUUCCAUGAUAUCU





1789
UUGCGUUCCAUGAUAUCUC





1790
UGCGUUCCAUGAUAUCUCA





1791
GCGUUCCAUGAUAUCUCAC





1792
CGUUCCAUGAUAUCUCACC





1793
GUUCCAUGAUAUCUCACCC





1794
UUCCAUGAUAUCUCACCCC





1795
UCCAUGAUAUCUCACCCCA





1796
CCAUGAUAUCUCACCCCAA





1797
CAUGAUAUCUCACCCCAAG





1798
AUGAUAUCUCACCCCAAGC





1799
UGAUAUCUCACCCCAAGCU





1800
GAUAUCUCACCCCAAGCUC





1801
AUAUCUCACCCCAAGCUCC





1802
UAUCUCACCCCAAGCUCCA





1803
AUCUCACCCCAAGCUCCAA





1804
UCUCACCCCAAGCUCCAAC





1805
CUCACCCCAAGCUCCAACG





1806
UCACCCCAAGCUCCAACGC





1807
CACCCCAAGCUCCAACGCA





1808
ACCCCAAGCUCCAACGCAU





1809
CCCCAAGCUCCAACGCAUU





1810
CCCAAGCUCCAACGCAUUU





1811
CCAAGCUCCAACGCAUUUU





1812
CAAGCUCCAACGCAUUUUC





1813
AAGCUCCAACGCAUUUUCU





1814
AGCUCCAACGCAUUUUCUA





1815
GCUCCAACGCAUUUUCUAG





1816
CUCCAACGCAUUUUCUAGU





1817
UCCAACGCAUUUUCUAGUG





1818
CCAACGCAUUUUCUAGUGA





1819
CAACGCAUUUUCUAGUGAU





1820
AACGCAUUUUCUAGUGAUU





1821
ACGCAUUUUCUAGUGAUUC





1822
CGCAUUUUCUAGUGAUUCC





1823
GCAUUUUCUAGUGAUUCCU





1824
CAUUUUCUAGUGAUUCCUA





1825
AUUUUCUAGUGAUUCCUAA





1826
UUUUCUAGUGAUUCCUAAG





1827
UUUCUAGUGAUUCCUAAGA





1828
UUCUAGUGAUUCCUAAGAA





1829
UCUAGUGAUUCCUAAGAAG





1830
CUAGUGAUUCCUAAGAAGC





1831
UAGUGAUUCCUAAGAAGCC





1832
AGUGAUUCCUAAGAAGCCA





1833
GUGAUUCCUAAGAAGCCAA





1834
UGAUUCCUAAGAAGCCAAU





1835
GAUUCCUAAGAAGCCAAUU





1836
AUUCCUAAGAAGCCAAUUG





1837
UUCCUAAGAAGCCAAUUGU





1838
UCCUAAGAAGCCAAUUGUC





1839
CCUAAGAAGCCAAUUGUCA





1840
CUAAGAAGCCAAUUGUCAG





1841
UAAGAAGCCAAUUGUCAGG





1842
AAGAAGCCAAUUGUCAGGU





1843
AGAAGCCAAUUGUCAGGUU





1844
GAAGCCAAUUGUCAGGUUA





1845
AAGCCAAUUGUCAGGUUAU





1846
AGCCAAUUGUCAGGUUAUC





1847
GCCAAUUGUCAGGUUAUCU





1848
CCAAUUGUCAGGUUAUCUG





1849
CAAUUGUCAGGUUAUCUGA





1850
AAUUGUCAGGUUAUCUGAA





1851
AUUGUCAGGUUAUCUGAAG





1852
UUGUCAGGUUAUCUGAAGC





1853
UGUCAGGUUAUCUGAAGCA





1854
GUCAGGUUAUCUGAAGCAG





1855
UCAGGUUAUCUGAAGCAGA





1856
CAGGUUAUCUGAAGCAGAA





1857
AGGUUAUCUGAAGCAGAAG





1858
GGUUAUCUGAAGCAGAAGA





1859
GUUAUCUGAAGCAGAAGAU





1860
UUAUCUGAAGCAGAAGAUU





1861
UAUCUGAAGCAGAAGAUUC





1862
AUCUGAAGCAGAAGAUUCU





1863
UCUGAAGCAGAAGAUUCUG





1864
CUGAAGCAGAAGAUUCUGA





1865
UGAAGCAGAAGAUUCUGAU





1866
GAAGCAGAAGAUUCUGAUG





1867
AAGCAGAAGAUUCUGAUGA





1868
AGCAGAAGAUUCUGAUGAA





1869
GCAGAAGAUUCUGAUGAAU





1870
CAGAAGAUUCUGAUGAAUC





1871
AGAAGAUUCUGAUGAAUCU





1872
GAAGAUUCUGAUGAAUCUC





1873
AAGAUUCUGAUGAAUCUCU





1874
AGAUUCUGAUGAAUCUCUU





1875
GAUUCUGAUGAAUCUCUUC





1876
AUUCUGAUGAAUCUCUUCU





1877
UUCUGAUGAAUCUCUUCUG





1878
UCUGAUGAAUCUCUUCUGG





1879
CUGAUGAAUCUCUUCUGGG





1880
UGAUGAAUCUCUUCUGGGG





1881
GAUGAAUCUCUUCUGGGGC





1882
AUGAAUCUCUUCUGGGGCA





1883
UGAAUCUCUUCUGGGGCAU





1884
GAAUCUCUUCUGGGGCAUU





1885
AAUCUCUUCUGGGGCAUUU





1886
AUCUCUUCUGGGGCAUUUA





1887
UCUCUUCUGGGGCAUUUAA





1888
CUCUUCUGGGGCAUUUAAU





1889
UCUUCUGGGGCAUUUAAUG





1890
CUUCUGGGGCAUUUAAUGA





1891
UUCUGGGGCAUUUAAUGAU





1892
UCUGGGGCAUUUAAUGAUU





1893
CUGGGGCAUUUAAUGAUUG





1894
UGGGGCAUUUAAUGAUUGU





1895
GGGGCAUUUAAUGAUUGUU





1896
GGGCAUUUAAUGAUUGUUG





1897
GGCAUUUAAUGAUUGUUGG





1898
GCAUUUAAUGAUUGUUGGC





1899
CAUUUAAUGAUUGUUGGCA





1900
AUUUAAUGAUUGUUGGCAA





1901
UUUAAUGAUUGUUGGCAAG





1902
UUAAUGAUUGUUGGCAAGA





1903
UAAUGAUUGUUGGCAAGAA





1904
AAUGAUUGUUGGCAAGAAG





1905
AUGAUUGUUGGCAAGAAGU





1906
UGAUUGUUGGCAAGAAGUG





1907
GAUUGUUGGCAAGAAGUGU





1908
AUUGUUGGCAAGAAGUGUG





1909
UUGUUGGCAAGAAGUGUGC





1910
UGUUGGCAAGAAGUGUGCU





1911
GUUGGCAAGAAGUGUGCUG





1912
UUGGCAAGAAGUGUGCUGC





1913
UGGCAAGAAGUGUGCUGCU





1914
GGCAAGAAGUGUGCUGCUA





1915
GCAAGAAGUGUGCUGCUAA





1916
CAAGAAGUGUGCUGCUAAC





1917
AAGAAGUGUGCUGCUAACC





1918
AGAAGUGUGCUGCUAACCU





1919
GAAGUGUGCUGCUAACCUG





1920
AAGUGUGCUGCUAACCUGG





1921
AGUGUGCUGCUAACCUGGG





1922
GUGUGCUGCUAACCUGGGC





1923
UGUGCUGCUAACCUGGGCC





1924
GUGCUGCUAACCUGGGCCU





1925
UGCUGCUAACCUGGGCCUG





1926
GCUGCUAACCUGGGCCUGA





1927
CUGCUAACCUGGGCCUGAC





1928
UGCUAACCUGGGCCUGACC





1929
GCUAACCUGGGCCUGACCA





1930
CUAACCUGGGCCUGACCAA





1931
UAACCUGGGCCUGACCAAU





1932
AACCUGGGCCUGACCAAUG





1933
ACCUGGGCCUGACCAAUGG





1934
CCUGGGCCUGACCAAUGGA





1935
CUGGGCCUGACCAAUGGAU





1936
UGGGCCUGACCAAUGGAUU





1937
GGGCCUGACCAAUGGAUUC





1938
GGCCUGACCAAUGGAUUCC





1939
GCCUGACCAAUGGAUUCCG





1940
CCUGACCAAUGGAUUCCGG





1941
CUGACCAAUGGAUUCCGGA





1942
UGACCAAUGGAUUCCGGAU





1943
GACCAAUGGAUUCCGGAUG





1944
ACCAAUGGAUUCCGGAUGG





1945
CCAAUGGAUUCCGGAUGGU





1946
CAAUGGAUUCCGGAUGGUU





1947
AAUGGAUUCCGGAUGGUUU





1948
AUGGAUUCCGGAUGGUUUU





1949
UGGAUUCCGGAUGGUUUUG





1950
GGAUUCCGGAUGGUUUUGA





1951
GAUUCCGGAUGGUUUUGAA





1952
AUUCCGGAUGGUUUUGAAU





1953
UUCCGGAUGGUUUUGAAUG





1954
UCCGGAUGGUUUUGAAUGA





1955
CCGGAUGGUUUUGAAUGAA





1956
CGGAUGGUUUUGAAUGAAG





1957
GGAUGGUUUUGAAUGAAGG





1958
GAUGGUUUUGAAUGAAGGG





1959
AUGGUUUUGAAUGAAGGGC





1960
UGGUUUUGAAUGAAGGGCC





1961
GGUUUUGAAUGAAGGGCCU





1962
GUUUUGAAUGAAGGGCCUG





1963
UUUUGAAUGAAGGGCCUGA





1964
UUUGAAUGAAGGGCCUGAG





1965
UUGAAUGAAGGGCCUGAGG





1966
UGAAUGAAGGGCCUGAGGG





1967
GAAUGAAGGGCCUGAGGGU





1968
AAUGAAGGGCCUGAGGGUG





1969
AUGAAGGGCCUGAGGGUGG





1970
UGAAGGGCCUGAGGGUGGG





1971
GAAGGGCCUGAGGGUGGGC





1972
AAGGGCCUGAGGGUGGGCA





1973
AGGGCCUGAGGGUGGGCAG





1974
GGGCCUGAGGGUGGGCAGU





1975
GGCCUGAGGGUGGGCAGUC





1976
GCCUGAGGGUGGGCAGUCU





1977
CCUGAGGGUGGGCAGUCUG





1978
CUGAGGGUGGGCAGUCUGU





1979
UGAGGGUGGGCAGUCUGUC





1980
GAGGGUGGGCAGUCUGUCU





1981
AGGGUGGGCAGUCUGUCUA





1982
GGGUGGGCAGUCUGUCUAU





1983
GGUGGGCAGUCUGUCUAUC





1984
GUGGGCAGUCUGUCUAUCA





1985
UGGGCAGUCUGUCUAUCAU





1986
GGGCAGUCUGUCUAUCAUG





1987
GGCAGUCUGUCUAUCAUGU





1988
GCAGUCUGUCUAUCAUGUA





1989
CAGUCUGUCUAUCAUGUAC





1990
AGUCUGUCUAUCAUGUACA





1991
GUCUGUCUAUCAUGUACAU





1992
UCUGUCUAUCAUGUACAUC





1993
CUGUCUAUCAUGUACAUCU





1994
UGUCUAUCAUGUACAUCUC





1995
GUCUAUCAUGUACAUCUCC





1996
UCUAUCAUGUACAUCUCCA





1997
CUAUCAUGUACAUCUCCAU





1998
UAUCAUGUACAUCUCCAUA





1999
AUCAUGUACAUCUCCAUAU





2000
UCAUGUACAUCUCCAUAUU





2001
CAUGUACAUCUCCAUAUUC





2002
AUGUACAUCUCCAUAUUCU





2003
UGUACAUCUCCAUAUUCUG





2004
GUACAUCUCCAUAUUCUGG





2005
UACAUCUCCAUAUUCUGGG





2006
ACAUCUCCAUAUUCUGGGA





2007
CAUCUCCAUAUUCUGGGAG





2008
AUCUCCAUAUUCUGGGAGG





2009
UCUCCAUAUUCUGGGAGGU





2010
CUCCAUAUUCUGGGAGGUC





2011
UCCAUAUUCUGGGAGGUCG





2012
CCAUAUUCUGGGAGGUCGU





2013
CAUAUUCUGGGAGGUCGUC





2014
AUAUUCUGGGAGGUCGUCA





2015
UAUUCUGGGAGGUCGUCAG





2016
AUUCUGGGAGGUCGUCAGU





2017
UUCUGGGAGGUCGUCAGUU





2018
UCUGGGAGGUCGUCAGUUG





2019
CUGGGAGGUCGUCAGUUGG





2020
UGGGAGGUCGUCAGUUGGG





2021
GGGAGGUCGUCAGUUGGGC





2022
GGAGGUCGUCAGUUGGGCU





2023
GAGGUCGUCAGUUGGGCUG





2024
AGGUCGUCAGUUGGGCUGG





2025
GGUCGUCAGUUGGGCUGGC





2026
GUCGUCAGUUGGGCUGGCC





2027
UCGUCAGUUGGGCUGGCCU





2028
CGUCAGUUGGGCUGGCCUC





2029
GUCAGUUGGGCUGGCCUCC





2030
UCAGUUGGGCUGGCCUCCU





2031
CAGUUGGGCUGGCCUCCUG





2032
AGUUGGGCUGGCCUCCUGG





2033
GUUGGGCUGGCCUCCUGGC





2034
UUGGGCUGGCCUCCUGGCU





2035
UGGGCUGGCCUCCUGGCUA





2036
GGGCUGGCCUCCUGGCUAA





2037
GGCUGGCCUCCUGGCUAAG





2038
GCUGGCCUCCUGGCUAAGA





2039
CUGGCCUCCUGGCUAAGAU





2040
UGGCCUCCUGGCUAAGAUU





2041
GGCCUCCUGGCUAAGAUUU





2042
GCCUCCUGGCUAAGAUUUU





2043
CCUCCUGGCUAAGAUUUUU





2044
CUCCUGGCUAAGAUUUUUG





2045
UCCUGGCUAAGAUUUUUGC





2046
CCUGGCUAAGAUUUUUGCA





2047
CUGGCUAAGAUUUUUGCAC





2048
UGGCUAAGAUUUUUGCACC





2049
GGCUAAGAUUUUUGCACCA





2050
GCUAAGAUUUUUGCACCAC





2051
CUAAGAUUUUUGCACCACA





2052
UAAGAUUUUUGCACCACAA





2053
AAGAUUUUUGCACCACAAG





2054
AGAUUUUUGCACCACAAGA





2055
GAUUUUUGCACCACAAGAG





2056
AUUUUUGCACCACAAGAGA





2057
UUUUUGCACCACAAGAGAU





2058
UUUUGCACCACAAGAGAUG





2059
UUUGCACCACAAGAGAUGC





2060
UUGCACCACAAGAGAUGCU





2061
UGCACCACAAGAGAUGCUG





2062
GCACCACAAGAGAUGCUGC





2063
CACCACAAGAGAUGCUGCA





2064
ACCACAAGAGAUGCUGCAU





2065
CCACAAGAGAUGCUGCAUG





2066
CACAAGAGAUGCUGCAUGU





2067
ACAAGAGAUGCUGCAUGUG





2068
CAAGAGAUGCUGCAUGUGU





2069
AAGAGAUGCUGCAUGUGUA





2070
AGAGAUGCUGCAUGUGUAC





2071
GAGAUGCUGCAUGUGUACA





2072
AGAUGCUGCAUGUGUACAA





2073
GAUGCUGCAUGUGUACAAA





2074
AUGCUGCAUGUGUACAAAU





2075
UGCUGCAUGUGUACAAAUC





2076
GCUGCAUGUGUACAAAUCA





2077
CUGCAUGUGUACAAAUCAC





2078
UGCAUGUGUACAAAUCACU





2079
GCAUGUGUACAAAUCACUA





2080
CAUGUGUACAAAUCACUAG





2081
AUGUGUACAAAUCACUAGC





2082
UGUGUACAAAUCACUAGCA





2083
GUGUACAAAUCACUAGCAA





2084
UGUACAAAUCACUAGCAAA





2085
GUACAAAUCACUAGCAAAU





2086
UACAAAUCACUAGCAAAUA





2087
ACAAAUCACUAGCAAAUAG





2088
CAAAUCACUAGCAAAUAGA





2089
AAAUCACUAGCAAAUAGAU





2090
AAUCACUAGCAAAUAGAUU





2091
AUCACUAGCAAAUAGAUUU





2092
UCACUAGCAAAUAGAUUUG





2093
CACUAGCAAAUAGAUUUGU





2094
ACUAGCAAAUAGAUUUGUU





2095
CUAGCAAAUAGAUUUGUUU





2096
UAGCAAAUAGAUUUGUUUC





2097
AGCAAAUAGAUUUGUUUCC





2098
GCAAAUAGAUUUGUUUCCC





2099
CAAAUAGAUUUGUUUCCCA





2100
AAAUAGAUUUGUUUCCCAU





2101
AAUAGAUUUGUUUCCCAUC





2102
AUAGAUUUGUUUCCCAUCA





2103
UAGAUUUGUUUCCCAUCAA





2104
AGAUUUGUUUCCCAUCAAC





2105
GAUUUGUUUCCCAUCAACU





2106
AUUUGUUUCCCAUCAACUU





2107
UUUGUUUCCCAUCAACUUA





2108
UUGUUUCCCAUCAACUUAG





2109
UGUUUCCCAUCAACUUAGC





2110
GUUUCCCAUCAACUUAGCC





2111
UUUCCCAUCAACUUAGCCA





2112
UUCCCAUCAACUUAGCCAC





2113
UCCCAUCAACUUAGCCACU





2114
CCCAUCAACUUAGCCACUG





2115
CCAUCAACUUAGCCACUGU





2116
CAUCAACUUAGCCACUGUU





2117
AUCAACUUAGCCACUGUUA





2118
UCAACUUAGCCACUGUUAA





2119
CAACUUAGCCACUGUUAAU





2120
AACUUAGCCACUGUUAAUG





2121
ACUUAGCCACUGUUAAUGU





2122
CUUAGCCACUGUUAAUGUA





2123
UUAGCCACUGUUAAUGUAA





2124
UAGCCACUGUUAAUGUAAA





2125
AGCCACUGUUAAUGUAAAU





2126
GCCACUGUUAAUGUAAAUU





2127
CCACUGUUAAUGUAAAUUG





2128
CACUGUUAAUGUAAAUUGU





2129
ACUGUUAAUGUAAAUUGUU





2130
CUGUUAAUGUAAAUUGUUC





2131
UGUUAAUGUAAAUUGUUCU





2132
GUUAAUGUAAAUUGUUCUU





2133
UUAAUGUAAAUUGUUCUUG





2134
UAAUGUAAAUUGUUCUUGG





2135
AAUGUAAAUUGUUCUUGGA





2136
AUGUAAAUUGUUCUUGGAU





2137
UGUAAAUUGUUCUUGGAUA





2138
GUAAAUUGUUCUUGGAUAU





2139
UAAAUUGUUCUUGGAUAUG





2140
AAAUUGUUCUUGGAUAUGU





2141
AAUUGUUCUUGGAUAUGUG





2142
AUUGUUCUUGGAUAUGUGU





2143
UUGUUCUUGGAUAUGUGUC





2144
UGUUCUUGGAUAUGUGUCU





2145
GUUCUUGGAUAUGUGUCUU





2146
UUCUUGGAUAUGUGUCUUU





2147
UCUUGGAUAUGUGUCUUUG





2148
CUUGGAUAUGUGUCUUUGG





2149
UUGGAUAUGUGUCUUUGGA





2150
UGGAUAUGUGUCUUUGGAG





2151
GGAUAUGUGUCUUUGGAGG





2152
GAUAUGUGUCUUUGGAGGG





2153
AUAUGUGUCUUUGGAGGGC





2154
UAUGUGUCUUUGGAGGGCA





2155
AUGUGUCUUUGGAGGGCAA





2156
UGUGUCUUUGGAGGGCAAU





2157
GUGUCUUUGGAGGGCAAUA





2158
UGUCUUUGGAGGGCAAUAA





2159
GUCUUUGGAGGGCAAUAAA





2160
UCUUUGGAGGGCAAUAAAU





2161
CUUUGGAGGGCAAUAAAUG





2162
UUUGGAGGGCAAUAAAUGC





2163
UUGGAGGGCAAUAAAUGCU





2164
UGGAGGGCAAUAAAUGCUC





2165
GGAGGGCAAUAAAUGCUCU





2166
GAGGGCAAUAAAUGCUCUG





2167
AGGGCAAUAAAUGCUCUGA





2168
GGGCAAUAAAUGCUCUGAA





2169
GGCAAUAAAUGCUCUGAAC





2170
GCAAUAAAUGCUCUGAACA





2171
CAAUAAAUGCUCUGAACAG





2172
AAUAAAUGCUCUGAACAGC





2173
AUAAAUGCUCUGAACAGCA





2174
UAAAUGCUCUGAACAGCAC





2175
AAAUGCUCUGAACAGCACU





2176
AAUGCUCUGAACAGCACUU





2177
AUGCUCUGAACAGCACUUG





2178
UGCUCUGAACAGCACUUGC





2179
GCUCUGAACAGCACUUGCA





2180
CUCUGAACAGCACUUGCAC





2181
UCUGAACAGCACUUGCACA





2182
CUGAACAGCACUUGCACAA





2183
UGAACAGCACUUGCACAAU





2184
GAACAGCACUUGCACAAUA





2185
CACUUGCACAAUAAAGAUA





2186
ACUUGCACAAUAAAGAUAC





2187
CUUGCACAAUAAAGAUACA





2188
UUGCACAAUAAAGAUACAG





2189
UGCACAAUAAAGAUACAGC





2190
GCACAAUAAAGAUACAGCA





2191
CACAAUAAAGAUACAGCAU





2192
ACAAUAAAGAUACAGCAUG





2193
CAAUAAAGAUACAGCAUGU





2194
AAUAAAGAUACAGCAUGUG





2195
AUAAAGAUACAGCAUGUGG





2196
UAAAGAUACAGCAUGUGGA





2197
AAAGAUACAGCAUGUGGAA





2198
AAGAUACAGCAUGUGGAAA





2199
AGAUACAGCAUGUGGAAAA





2200
GAUACAGCAUGUGGAAAAA





2201
AUACAGCAUGUGGAAAAAA





2202
UACAGCAUGUGGAAAAAAA





2203
ACAGCAUGUGGAAAAAAAA





2204
CAGCAUGUGGAAAAAAAAA





2205
AGCAUGUGGAAAAAAAAAA





2206
GCAUGUGGAAAAAAAAAAA





2207
CAUGUGGAAAAAAAAAAAA





2208
AUGUGGAAAAAAAAAAAAA





2209
UGUGGAAAAAAAAAAAAAA
















TABLE 3







DsRNA molecules targeting mRNA encoding chicken


r-spondin.








SEQ ID NO
Sequence 5′-3′





2210
AUGGAUCUAACAGGCGGCA





2211
UGGAUCUAACAGGCGGCAG





2212
GGAUCUAACAGGCGGCAGC





2213
GAUCUAACAGGCGGCAGCA





2214
AUCUAACAGGCGGCAGCAA





2215
UCUAACAGGCGGCAGCAAA





2216
CUAACAGGCGGCAGCAAAG





2217
UAACAGGCGGCAGCAAAGU





2218
AACAGGCGGCAGCAAAGUG





2219
ACAGGCGGCAGCAAAGUGG





2220
CAGGCGGCAGCAAAGUGGU





2221
AGGCGGCAGCAAAGUGGUG





2222
GGCGGCAGCAAAGUGGUGA





2223
GCGGCAGCAAAGUGGUGAA





2224
CGGCAGCAAAGUGGUGAAG





2225
GGCAGCAAAGUGGUGAAGG





2226
GCAGCAAAGUGGUGAAGGG





2227
CAGCAAAGUGGUGAAGGGC





2228
AGCAAAGUGGUGAAGGGCA





2229
GCAAAGUGGUGAAGGGCAA





2230
CAAAGUGGUGAAGGGCAAG





2231
AAAGUGGUGAAGGGCAAGA





2232
AAGUGGUGAAGGGCAAGAG





2233
AGUGGUGAAGGGCAAGAGG





2234
GUGGUGAAGGGCAAGAGGC





2235
UGGUGAAGGGCAAGAGGCA





2236
GGUGAAGGGCAAGAGGCAA





2237
GUGAAGGGCAAGAGGCAAA





2238
UGAAGGGCAAGAGGCAAAG





2239
GAAGGGCAAGAGGCAAAGG





2240
AAGGGCAAGAGGCAAAGGC





2241
AGGGCAAGAGGCAAAGGCG





2242
GGGCAAGAGGCAAAGGCGA





2243
GGCAAGAGGCAAAGGCGAA





2244
GCAAGAGGCAAAGGCGAAU





2245
CAAGAGGCAAAGGCGAAUU





2246
AAGAGGCAAAGGCGAAUUA





2247
AGAGGCAAAGGCGAAUUAG





2248
GAGGCAAAGGCGAAUUAGC





2249
AGGCAAAGGCGAAUUAGCA





2250
GGCAAAGGCGAAUUAGCAC





2251
GCAAAGGCGAAUUAGCACU





2252
CAAAGGCGAAUUAGCACUG





2253
AAAGGCGAAUUAGCACUGA





2254
AAGGCGAAUUAGCACUGAG





2255
AGGCGAAUUAGCACUGAGC





2256
GGCGAAUUAGCACUGAGCU





2257
GCGAAUUAGCACUGAGCUG





2258
CGAAUUAGCACUGAGCUGA





2259
GAAUUAGCACUGAGCUGAG





2260
AAUUAGCACUGAGCUGAGC





2261
AUUAGCACUGAGCUGAGCC





2262
UUAGCACUGAGCUGAGCCA





2263
UAGCACUGAGCUGAGCCAG





2264
AGCACUGAGCUGAGCCAGG





2265
GCACUGAGCUGAGCCAGGG





2266
CACUGAGCUGAGCCAGGGC





2267
ACUGAGCUGAGCCAGGGCU





2268
CUGAGCUGAGCCAGGGCUG





2269
UGAGCUGAGCCAGGGCUGU





2270
GAGCUGAGCCAGGGCUGUG





2271
AGCUGAGCCAGGGCUGUGC





2272
GCUGAGCCAGGGCUGUGCC





2273
CUGAGCCAGGGCUGUGCCA





2274
UGAGCCAGGGCUGUGCCAG





2275
GAGCCAGGGCUGUGCCAGG





2276
AGCCAGGGCUGUGCCAGGG





2277
GCCAGGGCUGUGCCAGGGG





2278
CCAGGGCUGUGCCAGGGGC





2279
CAGGGCUGUGCCAGGGGCU





2280
AGGGCUGUGCCAGGGGCUG





2281
GGGCUGUGCCAGGGGCUGC





2282
GGCUGUGCCAGGGGCUGCG





2283
GCUGUGCCAGGGGCUGCGA





2284
CUGUGCCAGGGGCUGCGAC





2285
UGUGCCAGGGGCUGCGACC





2286
GUGCCAGGGGCUGCGACCU





2287
UGCCAGGGGCUGCGACCUG





2288
GCCAGGGGCUGCGACCUGU





2289
CCAGGGGCUGCGACCUGUG





2290
CAGGGGCUGCGACCUGUGC





2291
AGGGGCUGCGACCUGUGCU





2292
GGGGCUGCGACCUGUGCUC





2293
GGGCUGCGACCUGUGCUCU





2294
GGCUGCGACCUGUGCUCUG





2295
GCUGCGACCUGUGCUCUGA





2296
CUGCGACCUGUGCUCUGAG





2297
UGCGACCUGUGCUCUGAGU





2298
GCGACCUGUGCUCUGAGUU





2299
CGACCUGUGCUCUGAGUUC





2300
GACCUGUGCUCUGAGUUCA





2301
ACCUGUGCUCUGAGUUCAA





2302
CCUGUGCUCUGAGUUCAAC





2303
CUGUGCUCUGAGUUCAACG





2304
UGUGCUCUGAGUUCAACGG





2305
GUGCUCUGAGUUCAACGGG





2306
UGCUCUGAGUUCAACGGGU





2307
GCUCUGAGUUCAACGGGUG





2308
CUCUGAGUUCAACGGGUGC





2309
UCUGAGUUCAACGGGUGCC





2310
CUGAGUUCAACGGGUGCCU





2311
UGAGUUCAACGGGUGCCUG





2312
GAGUUCAACGGGUGCCUGA





2313
AGUUCAACGGGUGCCUGAG





2314
GUUCAACGGGUGCCUGAGA





2315
UUCAACGGGUGCCUGAGAU





2316
UCAACGGGUGCCUGAGAUG





2317
CAACGGGUGCCUGAGAUGU





2318
AACGGGUGCCUGAGAUGUU





2319
ACGGGUGCCUGAGAUGUUC





2320
CGGGUGCCUGAGAUGUUCC





2321
GGGUGCCUGAGAUGUUCCC





2322
GGUGCCUGAGAUGUUCCCC





2323
GUGCCUGAGAUGUUCCCCC





2324
UGCCUGAGAUGUUCCCCCA





2325
GCCUGAGAUGUUCCCCCAA





2326
CCUGAGAUGUUCCCCCAAG





2327
CUGAGAUGUUCCCCCAAGC





2328
UGAGAUGUUCCCCCAAGCU





2329
GAGAUGUUCCCCCAAGCUC





2330
AGAUGUUCCCCCAAGCUCU





2331
GAUGUUCCCCCAAGCUCUU





2332
AUGUUCCCCCAAGCUCUUC





2333
UGUUCCCCCAAGCUCUUCA





2334
GUUCCCCCAAGCUCUUCAU





2335
UUCCCCCAAGCUCUUCAUC





2336
UCCCCCAAGCUCUUCAUCC





2337
CCCCCAAGCUCUUCAUCCU





2338
CCCCAAGCUCUUCAUCCUU





2339
CCCAAGCUCUUCAUCCUUC





2340
CCAAGCUCUUCAUCCUUCU





2341
CAAGCUCUUCAUCCUUCUG





2342
AAGCUCUUCAUCCUUCUGG





2343
AGCUCUUCAUCCUUCUGGA





2344
GCUCUUCAUCCUUCUGGAG





2345
CUCUUCAUCCUUCUGGAGA





2346
UCUUCAUCCUUCUGGAGAG





2347
CUUCAUCCUUCUGGAGAGG





2348
UUCAUCCUUCUGGAGAGGA





2349
UCAUCCUUCUGGAGAGGAA





2350
CAUCCUUCUGGAGAGGAAC





2351
AUCCUUCUGGAGAGGAACG





2352
UCCUUCUGGAGAGGAACGA





2353
CCUUCUGGAGAGGAACGAU





2354
CUUCUGGAGAGGAACGAUA





2355
UUCUGGAGAGGAACGAUAU





2356
UCUGGAGAGGAACGAUAUC





2357
CUGGAGAGGAACGAUAUCC





2358
UGGAGAGGAACGAUAUCCG





2359
GGAGAGGAACGAUAUCCGG





2360
GAGAGGAACGAUAUCCGGC





2361
AGAGGAACGAUAUCCGGCA





2362
GAGGAACGAUAUCCGGCAA





2363
AGGAACGAUAUCCGGCAAA





2364
GGAACGAUAUCCGGCAAAU





2365
GAACGAUAUCCGGCAAAUU





2366
AACGAUAUCCGGCAAAUUG





2367
ACGAUAUCCGGCAAAUUGG





2368
CGAUAUCCGGCAAAUUGGG





2369
GAUAUCCGGCAAAUUGGGA





2370
AUAUCCGGCAAAUUGGGAU





2371
UAUCCGGCAAAUUGGGAUC





2372
AUCCGGCAAAUUGGGAUCU





2373
UCCGGCAAAUUGGGAUCUG





2374
CCGGCAAAUUGGGAUCUGC





2375
CGGCAAAUUGGGAUCUGCC





2376
GGCAAAUUGGGAUCUGCCU





2377
GCAAAUUGGGAUCUGCCUC





2378
CAAAUUGGGAUCUGCCUCC





2379
AAAUUGGGAUCUGCCUCCC





2380
AAUUGGGAUCUGCCUCCCA





2381
AUUGGGAUCUGCCUCCCAU





2382
UUGGGAUCUGCCUCCCAUC





2383
UGGGAUCUGCCUCCCAUCC





2384
GGGAUCUGCCUCCCAUCCU





2385
GGAUCUGCCUCCCAUCCUG





2386
GAUCUGCCUCCCAUCCUGU





2387
AUCUGCCUCCCAUCCUGUC





2388
UCUGCCUCCCAUCCUGUCC





2389
CUGCCUCCCAUCCUGUCCA





2390
UGCCUCCCAUCCUGUCCAC





2391
GCCUCCCAUCCUGUCCACU





2392
CCUCCCAUCCUGUCCACUG





2393
CUCCCAUCCUGUCCACUGG





2394
UCCCAUCCUGUCCACUGGG





2395
CCCAUCCUGUCCACUGGGA





2396
CCAUCCUGUCCACUGGGAU





2397
CAUCCUGUCCACUGGGAUA





2398
AUCCUGUCCACUGGGAUAC





2399
UCCUGUCCACUGGGAUACU





2400
CCUGUCCACUGGGAUACUU





2401
CUGUCCACUGGGAUACUUU





2402
UGUCCACUGGGAUACUUUG





2403
GUCCACUGGGAUACUUUGG





2404
UCCACUGGGAUACUUUGGC





2405
CCACUGGGAUACUUUGGCC





2406
CACUGGGAUACUUUGGCCU





2407
ACUGGGAUACUUUGGCCUU





2408
CUGGGAUACUUUGGCCUUC





2409
UGGGAUACUUUGGCCUUCG





2410
GGGAUACUUUGGCCUUCGC





2411
GGAUACUUUGGCCUUCGCA





2412
GAUACUUUGGCCUUCGCAA





2413
AUACUUUGGCCUUCGCAAU





2414
UACUUUGGCCUUCGCAAUA





2415
ACUUUGGCCUUCGCAAUAC





2416
CUUUGGCCUUCGCAAUACA





2417
UUUGGCCUUCGCAAUACAG





2418
UUGGCCUUCGCAAUACAGA





2419
UGGCCUUCGCAAUACAGAC





2420
GGCCUUCGCAAUACAGACA





2421
GCCUUCGCAAUACAGACAU





2422
CCUUCGCAAUACAGACAUG





2423
CUUCGCAAUACAGACAUGA





2424
UUCGCAAUACAGACAUGAA





2425
UCGCAAUACAGACAUGAAC





2426
CGCAAUACAGACAUGAACA





2427
GCAAUACAGACAUGAACAA





2428
CAAUACAGACAUGAACAAG





2429
AAUACAGACAUGAACAAGU





2430
AUACAGACAUGAACAAGUG





2431
UACAGACAUGAACAAGUGC





2432
ACAGACAUGAACAAGUGCA





2433
CAGACAUGAACAAGUGCAU





2434
AGACAUGAACAAGUGCAUC





2435
GACAUGAACAAGUGCAUCA





2436
ACAUGAACAAGUGCAUCAA





2437
CAUGAACAAGUGCAUCAAA





2438
AUGAACAAGUGCAUCAAAU





2439
UGAACAAGUGCAUCAAAUG





2440
GAACAAGUGCAUCAAAUGC





2441
AACAAGUGCAUCAAAUGCA





2442
ACAAGUGCAUCAAAUGCAA





2443
CAAGUGCAUCAAAUGCAAA





2444
AAGUGCAUCAAAUGCAAAA





2445
AGUGCAUCAAAUGCAAAAU





2446
GUGCAUCAAAUGCAAAAUC





2447
UGCAUCAAAUGCAAAAUCG





2448
GCAUCAAAUGCAAAAUCGA





2449
CAUCAAAUGCAAAAUCGAG





2450
AUCAAAUGCAAAAUCGAGA





2451
UCAAAUGCAAAAUCGAGAA





2452
CAAAUGCAAAAUCGAGAAC





2453
AAAUGCAAAAUCGAGAACU





2454
AAUGCAAAAUCGAGAACUG





2455
AUGCAAAAUCGAGAACUGU





2456
UGCAAAAUCGAGAACUGUG





2457
GCAAAAUCGAGAACUGUGA





2458
CAAAAUCGAGAACUGUGAG





2459
AAAAUCGAGAACUGUGAGU





2460
AAAUCGAGAACUGUGAGUC





2461
AAUCGAGAACUGUGAGUCC





2462
AUCGAGAACUGUGAGUCCU





2463
UCGAGAACUGUGAGUCCUG





2464
CGAGAACUGUGAGUCCUGC





2465
GAGAACUGUGAGUCCUGCU





2466
AGAACUGUGAGUCCUGCUU





2467
GAACUGUGAGUCCUGCUUC





2468
AACUGUGAGUCCUGCUUCA





2469
ACUGUGAGUCCUGCUUCAG





2470
CUGUGAGUCCUGCUUCAGC





2471
UGUGAGUCCUGCUUCAGCC





2472
GUGAGUCCUGCUUCAGCCG





2473
UGAGUCCUGCUUCAGCCGA





2474
GAGUCCUGCUUCAGCCGAA





2475
AGUCCUGCUUCAGCCGAAA





2476
GUCCUGCUUCAGCCGAAAC





2477
UCCUGCUUCAGCCGAAACU





2478
CCUGCUUCAGCCGAAACUU





2479
CUGCUUCAGCCGAAACUUU





2480
UGCUUCAGCCGAAACUUUU





2481
GCUUCAGCCGAAACUUUUG





2482
CUUCAGCCGAAACUUUUGC





2483
UUCAGCCGAAACUUUUGCA





2484
UCAGCCGAAACUUUUGCAC





2485
CAGCCGAAACUUUUGCACA





2486
AGCCGAAACUUUUGCACAA





2487
GCCGAAACUUUUGCACAAA





2488
CCGAAACUUUUGCACAAAA





2489
CGAAACUUUUGCACAAAAU





2490
GAAACUUUUGCACAAAAUG





2491
AAACUUUUGCACAAAAUGU





2492
AACUUUUGCACAAAAUGUA





2493
ACUUUUGCACAAAAUGUAA





2494
CUUUUGCACAAAAUGUAAG





2495
UUUUGCACAAAAUGUAAGG





2496
UUUGCACAAAAUGUAAGGA





2497
UUGCACAAAAUGUAAGGAA





2498
UGCACAAAAUGUAAGGAAG





2499
GCACAAAAUGUAAGGAAGG





2500
CACAAAAUGUAAGGAAGGU





2501
ACAAAAUGUAAGGAAGGUU





2502
CAAAAUGUAAGGAAGGUUU





2503
AAAAUGUAAGGAAGGUUUG





2504
AAAUGUAAGGAAGGUUUGU





2505
AAUGUAAGGAAGGUUUGUA





2506
AUGUAAGGAAGGUUUGUAU





2507
UGUAAGGAAGGUUUGUAUU





2508
GUAAGGAAGGUUUGUAUUU





2509
UAAGGAAGGUUUGUAUUUG





2510
AAGGAAGGUUUGUAUUUGC





2511
AGGAAGGUUUGUAUUUGCA





2512
GGAAGGUUUGUAUUUGCAC





2513
GAAGGUUUGUAUUUGCACA





2514
AAGGUUUGUAUUUGCACAA





2515
AGGUUUGUAUUUGCACAAA





2516
GGUUUGUAUUUGCACAAAG





2517
GUUUGUAUUUGCACAAAGG





2518
UUUGUAUUUGCACAAAGGG





2519
UUGUAUUUGCACAAAGGGA





2520
UGUAUUUGCACAAAGGGAG





2521
GUAUUUGCACAAAGGGAGA





2522
UAUUUGCACAAAGGGAGAU





2523
AUUUGCACAAAGGGAGAUG





2524
UUUGCACAAAGGGAGAUGU





2525
UUGCACAAAGGGAGAUGUU





2526
UGCACAAAGGGAGAUGUUA





2527
GCACAAAGGGAGAUGUUAC





2528
CACAAAGGGAGAUGUUACG





2529
ACAAAGGGAGAUGUUACGU





2530
CAAAGGGAGAUGUUACGUC





2531
AAAGGGAGAUGUUACGUCA





2532
AAGGGAGAUGUUACGUCAC





2533
AGGGAGAUGUUACGUCACG





2534
GGGAGAUGUUACGUCACGU





2535
GGAGAUGUUACGUCACGUG





2536
GAGAUGUUACGUCACGUGC





2537
AGAUGUUACGUCACGUGCC





2538
GAUGUUACGUCACGUGCCC





2539
AUGUUACGUCACGUGCCCC





2540
UGUUACGUCACGUGCCCCG





2541
GUUACGUCACGUGCCCCGA





2542
UUACGUCACGUGCCCCGAA





2543
UACGUCACGUGCCCCGAAG





2544
ACGUCACGUGCCCCGAAGG





2545
CGUCACGUGCCCCGAAGGC





2546
GUCACGUGCCCCGAAGGCU





2547
UCACGUGCCCCGAAGGCUA





2548
CACGUGCCCCGAAGGCUAC





2549
ACGUGCCCCGAAGGCUACU





2550
CGUGCCCCGAAGGCUACUC





2551
GUGCCCCGAAGGCUACUCU





2552
UGCCCCGAAGGCUACUCUG





2553
GCCCCGAAGGCUACUCUGC





2554
CCCCGAAGGCUACUCUGCU





2555
CCCGAAGGCUACUCUGCUG





2556
CCGAAGGCUACUCUGCUGC





2557
CGAAGGCUACUCUGCUGCC





2558
GAAGGCUACUCUGCUGCCA





2559
AAGGCUACUCUGCUGCCAA





2560
AGGCUACUCUGCUGCCAAU





2561
GGCUACUCUGCUGCCAAUG





2562
GCUACUCUGCUGCCAAUGG





2563
CUACUCUGCUGCCAAUGGC





2564
UACUCUGCUGCCAAUGGCA





2565
ACUCUGCUGCCAAUGGCAC





2566
CUCUGCUGCCAAUGGCACC





2567
UCUGCUGCCAAUGGCACCA





2568
CUGCUGCCAAUGGCACCAU





2569
UGCUGCCAAUGGCACCAUG





2570
GCUGCCAAUGGCACCAUGG





2571
CUGCCAAUGGCACCAUGGA





2572
UGCCAAUGGCACCAUGGAG





2573
GCCAAUGGCACCAUGGAGU





2574
CCAAUGGCACCAUGGAGUG





2575
CAAUGGCACCAUGGAGUGC





2576
AAUGGCACCAUGGAGUGCA





2577
AUGGCACCAUGGAGUGCAG





2578
UGGCACCAUGGAGUGCAGC





2579
GGCACCAUGGAGUGCAGCA





2580
GCACCAUGGAGUGCAGCAG





2581
CACCAUGGAGUGCAGCAGU





2582
ACCAUGGAGUGCAGCAGUC





2583
CCAUGGAGUGCAGCAGUCC





2584
CAUGGAGUGCAGCAGUCCU





2585
AUGGAGUGCAGCAGUCCUG





2586
UGGAGUGCAGCAGUCCUGC





2587
GGAGUGCAGCAGUCCUGCG





2588
GAGUGCAGCAGUCCUGCGC





2589
AGUGCAGCAGUCCUGCGCA





2590
GUGCAGCAGUCCUGCGCAA





2591
UGCAGCAGUCCUGCGCAAU





2592
GCAGCAGUCCUGCGCAAUG





2593
CAGCAGUCCUGCGCAAUGU





2594
AGCAGUCCUGCGCAAUGUG





2595
GCAGUCCUGCGCAAUGUGA





2596
CAGUCCUGCGCAAUGUGAA





2597
AGUCCUGCGCAAUGUGAAA





2598
GUCCUGCGCAAUGUGAAAU





2599
UCCUGCGCAAUGUGAAAUG





2600
CCUGCGCAAUGUGAAAUGA





2601
CUGCGCAAUGUGAAAUGAG





2602
UGCGCAAUGUGAAAUGAGU





2603
GCGCAAUGUGAAAUGAGUG





2604
CGCAAUGUGAAAUGAGUGA





2605
GCAAUGUGAAAUGAGUGAG





2606
CAAUGUGAAAUGAGUGAGU





2607
AAUGUGAAAUGAGUGAGUG





2608
AUGUGAAAUGAGUGAGUGG





2609
UGUGAAAUGAGUGAGUGGG





2610
GUGAAAUGAGUGAGUGGGG





2611
UGAAAUGAGUGAGUGGGGG





2612
GAAAUGAGUGAGUGGGGGC





2613
AAAUGAGUGAGUGGGGGCC





2614
AAUGAGUGAGUGGGGGCCC





2615
AUGAGUGAGUGGGGGCCCU





2616
UGAGUGAGUGGGGGCCCUG





2617
GAGUGAGUGGGGGCCCUGG





2618
AGUGAGUGGGGGCCCUGGG





2619
GUGAGUGGGGGCCCUGGGG





2620
UGAGUGGGGGCCCUGGGGG





2621
GAGUGGGGGCCCUGGGGGC





2622
AGUGGGGGCCCUGGGGGCC





2623
GUGGGGGCCCUGGGGGCCC





2624
UGGGGGCCCUGGGGGCCCU





2625
GGGGGCCCUGGGGGCCCUG





2626
GGGGCCCUGGGGGCCCUGC





2627
GGGCCCUGGGGGCCCUGCU





2628
GGCCCUGGGGGCCCUGCUC





2629
GCCCUGGGGGCCCUGCUCC





2630
CCCUGGGGGCCCUGCUCCA





2631
CCUGGGGGCCCUGCUCCAA





2632
CUGGGGGCCCUGCUCCAAG





2633
UGGGGGCCCUGCUCCAAGA





2634
GGGGGCCCUGCUCCAAGAA





2635
GGGGCCCUGCUCCAAGAAG





2636
GGGCCCUGCUCCAAGAAGA





2637
GGCCCUGCUCCAAGAAGAG





2638
GCCCUGCUCCAAGAAGAGG





2639
CCCUGCUCCAAGAAGAGGA





2640
CCUGCUCCAAGAAGAGGAA





2641
CUGCUCCAAGAAGAGGAAG





2642
UGCUCCAAGAAGAGGAAGC





2643
GCUCCAAGAAGAGGAAGCU





2644
CUCCAAGAAGAGGAAGCUG





2645
UCCAAGAAGAGGAAGCUGU





2646
CCAAGAAGAGGAAGCUGUG





2647
CAAGAAGAGGAAGCUGUGU





2648
AAGAAGAGGAAGCUGUGUG





2649
AGAAGAGGAAGCUGUGUGG





2650
GAAGAGGAAGCUGUGUGGC





2651
AAGAGGAAGCUGUGUGGCU





2652
AGAGGAAGCUGUGUGGCUU





2653
GAGGAAGCUGUGUGGCUUC





2654
AGGAAGCUGUGUGGCUUCA





2655
GGAAGCUGUGUGGCUUCAA





2656
GAAGCUGUGUGGCUUCAAG





2657
AAGCUGUGUGGCUUCAAGA





2658
AGCUGUGUGGCUUCAAGAA





2659
GCUGUGUGGCUUCAAGAAG





2660
CUGUGUGGCUUCAAGAAGG





2661
UGUGUGGCUUCAAGAAGGG





2662
GUGUGGCUUCAAGAAGGGG





2663
UGUGGCUUCAAGAAGGGGA





2664
GUGGCUUCAAGAAGGGGAA





2665
UGGCUUCAAGAAGGGGAAC





2666
GGCUUCAAGAAGGGGAACG





2667
GCUUCAAGAAGGGGAACGA





2668
CUUCAAGAAGGGGAACGAG





2669
UUCAAGAAGGGGAACGAGG





2670
UCAAGAAGGGGAACGAGGA





2671
CAAGAAGGGGAACGAGGAC





2672
AAGAAGGGGAACGAGGACC





2673
AGAAGGGGAACGAGGACCG





2674
GAAGGGGAACGAGGACCGA





2675
AAGGGGAACGAGGACCGAA





2676
AGGGGAACGAGGACCGAAC





2677
GGGGAACGAGGACCGAACG





2678
GGGAACGAGGACCGAACGC





2679
GGAACGAGGACCGAACGCG





2680
GAACGAGGACCGAACGCGG





2681
AACGAGGACCGAACGCGGC





2682
ACGAGGACCGAACGCGGCG





2683
CGAGGACCGAACGCGGCGG





2684
GAGGACCGAACGCGGCGGA





2685
AGGACCGAACGCGGCGGAU





2686
GGACCGAACGCGGCGGAUC





2687
GACCGAACGCGGCGGAUCC





2688
ACCGAACGCGGCGGAUCCU





2689
CCGAACGCGGCGGAUCCUG





2690
CGAACGCGGCGGAUCCUGC





2691
GAACGCGGCGGAUCCUGCA





2692
AACGCGGCGGAUCCUGCAG





2693
ACGCGGCGGAUCCUGCAGG





2694
CGCGGCGGAUCCUGCAGGC





2695
GCGGCGGAUCCUGCAGGCU





2696
CGGCGGAUCCUGCAGGCUC





2697
GGCGGAUCCUGCAGGCUCC





2698
GCGGAUCCUGCAGGCUCCC





2699
CGGAUCCUGCAGGCUCCCU





2700
GGAUCCUGCAGGCUCCCUC





2701
GAUCCUGCAGGCUCCCUCU





2702
AUCCUGCAGGCUCCCUCUG





2703
UCCUGCAGGCUCCCUCUGG





2704
CCUGCAGGCUCCCUCUGGG





2705
CUGCAGGCUCCCUCUGGGG





2706
UGCAGGCUCCCUCUGGGGA





2707
GCAGGCUCCCUCUGGGGAC





2708
CAGGCUCCCUCUGGGGACG





2709
AGGCUCCCUCUGGGGACGU





2710
GGCUCCCUCUGGGGACGUG





2711
GCUCCCUCUGGGGACGUGU





2712
CUCCCUCUGGGGACGUGUC





2713
UCCCUCUGGGGACGUGUCC





2714
CCCUCUGGGGACGUGUCCC





2715
CCUCUGGGGACGUGUCCCU





2716
CUCUGGGGACGUGUCCCUG





2717
UCUGGGGACGUGUCCCUGU





2718
CUGGGGACGUGUCCCUGUG





2719
UGGGGACGUGUCCCUGUGC





2720
GGGGACGUGUCCCUGUGCC





2721
GGGACGUGUCCCUGUGCCC





2722
GGACGUGUCCCUGUGCCCC





2723
GACGUGUCCCUGUGCCCCG





2724
ACGUGUCCCUGUGCCCCGC





2725
CGUGUCCCUGUGCCCCGCC





2726
GUGUCCCUGUGCCCCGCCA





2727
UGUCCCUGUGCCCCGCCAC





2728
GUCCCUGUGCCCCGCCACC





2729
UCCCUGUGCCCCGCCACCA





2730
CCCUGUGCCCCGCCACCAC





2731
CCUGUGCCCCGCCACCACG





2732
CUGUGCCCCGCCACCACGG





2733
UGUGCCCCGCCACCACGGA





2734
GUGCCCCGCCACCACGGAG





2735
UGCCCCGCCACCACGGAGG





2736
GCCCCGCCACCACGGAGGU





2737
CCCCGCCACCACGGAGGUG





2738
CCCGCCACCACGGAGGUGC





2739
CCGCCACCACGGAGGUGCG





2740
CGCCACCACGGAGGUGCGC





2741
GCCACCACGGAGGUGCGCA





2742
CCACCACGGAGGUGCGCAG





2743
CACCACGGAGGUGCGCAGA





2744
ACCACGGAGGUGCGCAGAU





2745
CCACGGAGGUGCGCAGAUG





2746
CACGGAGGUGCGCAGAUGC





2747
ACGGAGGUGCGCAGAUGCA





2748
CGGAGGUGCGCAGAUGCAC





2749
GGAGGUGCGCAGAUGCACU





2750
GAGGUGCGCAGAUGCACUG





2751
AGGUGCGCAGAUGCACUGU





2752
GGUGCGCAGAUGCACUGUG





2753
GUGCGCAGAUGCACUGUGC





2754
UGCGCAGAUGCACUGUGCA





2755
GCGCAGAUGCACUGUGCAG





2756
CGCAGAUGCACUGUGCAGA





2757
GCAGAUGCACUGUGCAGAA





2758
CAGAUGCACUGUGCAGAAG





2759
AGAUGCACUGUGCAGAAGA





2760
GAUGCACUGUGCAGAAGAG





2761
AUGCACUGUGCAGAAGAGC





2762
UGCACUGUGCAGAAGAGCC





2763
GCACUGUGCAGAAGAGCCA





2764
CACUGUGCAGAAGAGCCAA





2765
ACUGUGCAGAAGAGCCAAU





2766
CUGUGCAGAAGAGCCAAUG





2767
UGUGCAGAAGAGCCAAUGC





2768
GUGCAGAAGAGCCAAUGCC





2769
UGCAGAAGAGCCAAUGCCC





2770
GCAGAAGAGCCAAUGCCCC





2771
CAGAAGAGCCAAUGCCCCG





2772
AGAAGAGCCAAUGCCCCGA





2773
GAAGAGCCAAUGCCCCGAA





2774
AAGAGCCAAUGCCCCGAAG





2775
AGAGCCAAUGCCCCGAAGG





2776
GAGCCAAUGCCCCGAAGGG





2777
AGCCAAUGCCCCGAAGGGA





2778
GCCAAUGCCCCGAAGGGAA





2779
CCAAUGCCCCGAAGGGAAA





2780
CAAUGCCCCGAAGGGAAAA





2781
AAUGCCCCGAAGGGAAAAG





2782
AUGCCCCGAAGGGAAAAGG





2783
UGCCCCGAAGGGAAAAGGA





2784
GCCCCGAAGGGAAAAGGAA





2785
CCCCGAAGGGAAAAGGAAG





2786
CCCGAAGGGAAAAGGAAGA





2787
CCGAAGGGAAAAGGAAGAA





2788
CGAAGGGAAAAGGAAGAAA





2789
GAAGGGAAAAGGAAGAAAA





2790
AAGGGAAAAGGAAGAAAAA





2791
AGGGAAAAGGAAGAAAAAG





2792
GGGAAAAGGAAGAAAAAGG





2793
GGAAAAGGAAGAAAAAGGA





2794
GAAAAGGAAGAAAAAGGAC





2795
AAAAGGAAGAAAAAGGACG





2796
AAAGGAAGAAAAAGGACGA





2797
AAGGAAGAAAAAGGACGAG





2798
AGGAAGAAAAAGGACGAGC





2799
GGAAGAAAAAGGACGAGCA





2800
GAAGAAAAAGGACGAGCAA





2801
AAGAAAAAGGACGAGCAAG





2802
AGAAAAAGGACGAGCAAGG





2803
GAAAAAGGACGAGCAAGGA





2804
AAAAAGGACGAGCAAGGAA





2805
AAAAGGACGAGCAAGGAAA





2806
AAAGGACGAGCAAGGAAAG





2807
AAGGACGAGCAAGGAAAGC





2808
AGGACGAGCAAGGAAAGCA





2809
GGACGAGCAAGGAAAGCAA





2810
GACGAGCAAGGAAAGCAAG





2811
ACGAGCAAGGAAAGCAAGA





2812
CGAGCAAGGAAAGCAAGAU





2813
GAGCAAGGAAAGCAAGAUA





2814
AGCAAGGAAAGCAAGAUAA





2815
GCAAGGAAAGCAAGAUAAU





2816
CAAGGAAAGCAAGAUAAUA





2817
AAGGAAAGCAAGAUAAUAC





2818
AGGAAAGCAAGAUAAUACA





2819
GGAAAGCAAGAUAAUACAA





2820
GAAAGCAAGAUAAUACAAA





2821
AAAGCAAGAUAAUACAAAC





2822
AAGCAAGAUAAUACAAACG





2823
AGCAAGAUAAUACAAACGG





2824
GCAAGAUAAUACAAACGGG





2825
CAAGAUAAUACAAACGGGA





2826
AAGAUAAUACAAACGGGAA





2827
AGAUAAUACAAACGGGAAC





2828
GAUAAUACAAACGGGAACA





2829
AUAAUACAAACGGGAACAG





2830
UAAUACAAACGGGAACAGA





2831
AAUACAAACGGGAACAGAA





2832
AUACAAACGGGAACAGAAA





2833
UACAAACGGGAACAGAAAU





2834
ACAAACGGGAACAGAAAUC





2835
CAAACGGGAACAGAAAUCG





2836
AAACGGGAACAGAAAUCGG





2837
AACGGGAACAGAAAUCGGA





2838
ACGGGAACAGAAAUCGGAA





2839
CGGGAACAGAAAUCGGAAA





2840
GGGAACAGAAAUCGGAAAG





2841
GGAACAGAAAUCGGAAAGA





2842
GAACAGAAAUCGGAAAGAC





2843
AACAGAAAUCGGAAAGACA





2844
ACAGAAAUCGGAAAGACAC





2845
CAGAAAUCGGAAAGACACC





2846
AGAAAUCGGAAAGACACCA





2847
GAAAUCGGAAAGACACCAA





2848
AAAUCGGAAAGACACCAAA





2849
AAUCGGAAAGACACCAAAG





2850
AUCGGAAAGACACCAAAGA





2851
UCGGAAAGACACCAAAGAU





2852
CGGAAAGACACCAAAGAUG





2853
GGAAAGACACCAAAGAUGC





2854
GAAAGACACCAAAGAUGCA





2855
AAAGACACCAAAGAUGCAA





2856
AAGACACCAAAGAUGCAAA





2857
AGACACCAAAGAUGCAAAG





2858
GACACCAAAGAUGCAAAGU





2859
ACACCAAAGAUGCAAAGUC





2860
CACCAAAGAUGCAAAGUCU





2861
ACCAAAGAUGCAAAGUCUG





2862
CCAAAGAUGCAAAGUCUGG





2863
CAAAGAUGCAAAGUCUGGC





2864
AAAGAUGCAAAGUCUGGCA





2865
AAGAUGCAAAGUCUGGCAC





2866
AGAUGCAAAGUCUGGCACC





2867
GAUGCAAAGUCUGGCACCA





2868
AUGCAAAGUCUGGCACCAA





2869
UGCAAAGUCUGGCACCAAG





2870
GCAAAGUCUGGCACCAAGA





2871
CAAAGUCUGGCACCAAGAA





2872
AAAGUCUGGCACCAAGAAG





2873
AAGUCUGGCACCAAGAAGA





2874
AGUCUGGCACCAAGAAGAG





2875
GUCUGGCACCAAGAAGAGG





2876
UCUGGCACCAAGAAGAGGA





2877
CUGGCACCAAGAAGAGGAA





2878
UGGCACCAAGAAGAGGAAG





2879
GGCACCAAGAAGAGGAAGA





2880
GCACCAAGAAGAGGAAGAG





2881
CACCAAGAAGAGGAAGAGC





2882
ACCAAGAAGAGGAAGAGCA





2883
CCAAGAAGAGGAAGAGCAA





2884
CAAGAAGAGGAAGAGCAAA





2885
AAGAAGAGGAAGAGCAAAC





2886
AGAAGAGGAAGAGCAAACA





2887
GAAGAGGAAGAGCAAACAG





2888
AAGAGGAAGAGCAAACAGA





2889
AGAGGAAGAGCAAACAGAG





2890
GAGGAAGAGCAAACAGAGG





2891
AGGAAGAGCAAACAGAGGG





2892
GGAAGAGCAAACAGAGGGG





2893
GAAGAGCAAACAGAGGGGG





2894
AAGAGCAAACAGAGGGGGG





2895
AGAGCAAACAGAGGGGGGC





2896
GAGCAAACAGAGGGGGGCU





2897
AGCAAACAGAGGGGGGCUG





2898
GCAAACAGAGGGGGGCUGU





2899
CAAACAGAGGGGGGCUGUG





2900
AAACAGAGGGGGGCUGUGG





2901
AACAGAGGGGGGCUGUGGC





2902
ACAGAGGGGGGCUGUGGCC





2903
CAGAGGGGGGCUGUGGCCC





2904
AGAGGGGGGCUGUGGCCCC





2905
GAGGGGGGCUGUGGCCCCC





2906
AGGGGGGCUGUGGCCCCCA





2907
GGGGGGCUGUGGCCCCCAC





2908
GGGGGCUGUGGCCCCCACC





2909
GGGGCUGUGGCCCCCACCA





2910
GGGCUGUGGCCCCCACCAC





2911
GGCUGUGGCCCCCACCACA





2912
GCUGUGGCCCCCACCACAU





2913
CUGUGGCCCCCACCACAUC





2914
UGUGGCCCCCACCACAUCC





2915
GUGGCCCCCACCACAUCCG





2916
UGGCCCCCACCACAUCCGC





2917
GGCCCCCACCACAUCCGCC





2918
GCCCCCACCACAUCCGCCA





2919
CCCCCACCACAUCCGCCAG





2920
CCCCACCACAUCCGCCAGC





2921
CCCACCACAUCCGCCAGCC





2922
CCACCACAUCCGCCAGCCC





2923
CACCACAUCCGCCAGCCCU





2924
ACCACAUCCGCCAGCCCUG





2925
CCACAUCCGCCAGCCCUGC





2926
CACAUCCGCCAGCCCUGCC





2927
ACAUCCGCCAGCCCUGCCC





2928
CAUCCGCCAGCCCUGCCCA





2929
AUCCGCCAGCCCUGCCCAA





2930
UCCGCCAGCCCUGCCCAAU





2931
CCGCCAGCCCUGCCCAAUA





2932
CGCCAGCCCUGCCCAAUAG





2933
GCCAGCCCUGCCCAAUAGC





2934
CCAGCCCUGCCCAAUAGCU





2935
CAGCCCUGCCCAAUAGCUG





2936
AGCCCUGCCCAAUAGCUGC





2937
GCCCUGCCCAAUAGCUGCC





2938
CCCUGCCCAAUAGCUGCCC





2939
CCUGCCCAAUAGCUGCCCC





2940
CUGCCCAAUAGCUGCCCCU





2941
UGCCCAAUAGCUGCCCCUU





2942
GCCCAAUAGCUGCCCCUUU





2943
CCCAAUAGCUGCCCCUUUA





2944
CCAAUAGCUGCCCCUUUAC





2945
CAAUAGCUGCCCCUUUACG





2946
AAUAGCUGCCCCUUUACGU





2947
AUAGCUGCCCCUUUACGUC





2948
UAGCUGCCCCUUUACGUCA





2949
AGCUGCCCCUUUACGUCAC





2950
GCUGCCCCUUUACGUCACC





2951
CUGCCCCUUUACGUCACCU





2952
UGCCCCUUUACGUCACCUG





2953
GCCCCUUUACGUCACCUGA





2954
CCCCUUUACGUCACCUGAC





2955
CCCUUUACGUCACCUGACG





2956
CCUUUACGUCACCUGACGG





2957
CUUUACGUCACCUGACGGC





2958
UUUACGUCACCUGACGGCA





2959
UUACGUCACCUGACGGCAA





2960
UACGUCACCUGACGGCAAG





2961
ACGUCACCUGACGGCAAGA





2962
CGUCACCUGACGGCAAGAC





2963
GUCACCUGACGGCAAGACU





2964
UCACCUGACGGCAAGACUU





2965
CACCUGACGGCAAGACUUC





2966
ACCUGACGGCAAGACUUCA





2967
CCUGACGGCAAGACUUCAU





2968
CUGACGGCAAGACUUCAUU





2969
UGACGGCAAGACUUCAUUG





2970
GACGGCAAGACUUCAUUGC





2971
ACGGCAAGACUUCAUUGCU





2972
CGGCAAGACUUCAUUGCUG





2973
GGCAAGACUUCAUUGCUGC





2974
GCAAGACUUCAUUGCUGCU





2975
CAAGACUUCAUUGCUGCUA





2976
AAGACUUCAUUGCUGCUAU





2977
AGACUUCAUUGCUGCUAUG





2978
GACUUCAUUGCUGCUAUGU





2979
ACUUCAUUGCUGCUAUGUA





2980
CUUCAUUGCUGCUAUGUAU





2981
UUCAUUGCUGCUAUGUAUA





2982
UCAUUGCUGCUAUGUAUAU





2983
CAUUGCUGCUAUGUAUAUG





2984
AUUGCUGCUAUGUAUAUGA





2985
UUGCUGCUAUGUAUAUGAA





2986
UGCUGCUAUGUAUAUGAAA





2987
GCUGCUAUGUAUAUGAAAG





2988
CUGCUAUGUAUAUGAAAGC





2989
UGCUAUGUAUAUGAAAGCU





2990
GCUAUGUAUAUGAAAGCUU





2991
CUAUGUAUAUGAAAGCUUU





2992
UAUGUAUAUGAAAGCUUUA





2993
AUGUAUAUGAAAGCUUUAU





2994
UGUAUAUGAAAGCUUUAUU





2995
GUAUAUGAAAGCUUUAUUG





2996
UAUAUGAAAGCUUUAUUGA





2997
AUAUGAAAGCUUUAUUGAA





2998
UAUGAAAGCUUUAUUGAAC





2999
AUGAAAGCUUUAUUGAACC





3000
UGAAAGCUUUAUUGAACCA





3001
GAAAGCUUUAUUGAACCAG





3002
AAAGCUUUAUUGAACCAGA





3003
AAGCUUUAUUGAACCAGAG





3004
AGCUUUAUUGAACCAGAGC





3005
GCUUUAUUGAACCAGAGCA





3006
CUUUAUUGAACCAGAGCAC





3007
UUUAUUGAACCAGAGCACU





3008
UUAUUGAACCAGAGCACUG





3009
UAUUGAACCAGAGCACUGC





3010
AUUGAACCAGAGCACUGCU





3011
UUGAACCAGAGCACUGCUA





3012
UGAACCAGAGCACUGCUAC





3013
GAACCAGAGCACUGCUACA





3014
AACCAGAGCACUGCUACAC





3015
ACCAGAGCACUGCUACACA





3016
CCAGAGCACUGCUACACAA





3017
CAGAGCACUGCUACACAAC





3018
AGAGCACUGCUACACAACA





3019
GAGCACUGCUACACAACAU





3020
AGCACUGCUACACAACAUU





3021
GCACUGCUACACAACAUUA





3022
CACUGCUACACAACAUUAC





3023
ACUGCUACACAACAUUACA





3024
CUGCUACACAACAUUACAC





3025
UGCUACACAACAUUACACA





3026
GCUACACAACAUUACACAU





3027
CUACACAACAUUACACAUG





3028
UACACAACAUUACACAUGU





3029
ACACAACAUUACACAUGUC





3030
CACAACAUUACACAUGUCA





3031
ACAACAUUACACAUGUCAG





3032
CAACAUUACACAUGUCAGA





3033
AACAUUACACAUGUCAGAA





3034
ACAUUACACAUGUCAGAAA





3035
CAUUACACAUGUCAGAAAG





3036
AUUACACAUGUCAGAAAGA





3037
UUACACAUGUCAGAAAGAC





3038
UACACAUGUCAGAAAGACA





3039
ACACAUGUCAGAAAGACAG





3040
CACAUGUCAGAAAGACAGA





3041
ACAUGUCAGAAAGACAGAG





3042
CAUGUCAGAAAGACAGAGC





3043
AUGUCAGAAAGACAGAGCU





3044
UGUCAGAAAGACAGAGCUA





3045
GUCAGAAAGACAGAGCUAU





3046
UCAGAAAGACAGAGCUAUA





3047
CAGAAAGACAGAGCUAUAC





3048
AGAAAGACAGAGCUAUACU





3049
GAAAGACAGAGCUAUACUC





3050
AAAGACAGAGCUAUACUCC





3051
AAGACAGAGCUAUACUCCU





3052
AGACAGAGCUAUACUCCUA





3053
GACAGAGCUAUACUCCUAG





3054
ACAGAGCUAUACUCCUAGA





3055
CAGAGCUAUACUCCUAGAC





3056
AGAGCUAUACUCCUAGACU





3057
GAGCUAUACUCCUAGACUC





3058
AGCUAUACUCCUAGACUCG





3059
GCUAUACUCCUAGACUCGA





3060
CUAUACUCCUAGACUCGAC





3061
UAUACUCCUAGACUCGACA





3062
AUACUCCUAGACUCGACAG





3063
UACUCCUAGACUCGACAGA





3064
ACUCCUAGACUCGACAGAA





3065
CUCCUAGACUCGACAGAAG





3066
UCCUAGACUCGACAGAAGC





3067
CCUAGACUCGACAGAAGCC





3068
CUAGACUCGACAGAAGCCA





3069
UAGACUCGACAGAAGCCAC





3070
AGACUCGACAGAAGCCACA





3071
GACUCGACAGAAGCCACAU





3072
ACUCGACAGAAGCCACAUC





3073
CUCGACAGAAGCCACAUCC





3074
UCGACAGAAGCCACAUCCA





3075
CGACAGAAGCCACAUCCAC





3076
GACAGAAGCCACAUCCACA





3077
ACAGAAGCCACAUCCACAA





3078
CAGAAGCCACAUCCACAAC





3079
AGAAGCCACAUCCACAACA





3080
GAAGCCACAUCCACAACAC





3081
AAGCCACAUCCACAACACU





3082
AGCCACAUCCACAACACUU





3083
GCCACAUCCACAACACUUA





3084
CCACAUCCACAACACUUAA





3085
CACAUCCACAACACUUAAG





3086
ACAUCCACAACACUUAAGG





3087
CAUCCACAACACUUAAGGA





3088
AUCCACAACACUUAAGGAG





3089
UCCACAACACUUAAGGAGG





3090
CCACAACACUUAAGGAGGC





3091
CACAACACUUAAGGAGGCG





3092
ACAACACUUAAGGAGGCGG





3093
CAACACUUAAGGAGGCGGU





3094
AACACUUAAGGAGGCGGUA





3095
ACACUUAAGGAGGCGGUAC





3096
CACUUAAGGAGGCGGUACC





3097
ACUUAAGGAGGCGGUACCC





3098
CUUAAGGAGGCGGUACCCC





3099
UUAAGGAGGCGGUACCCCC





3100
UAAGGAGGCGGUACCCCCG





3101
AAGGAGGCGGUACCCCCGG





3102
AGGAGGCGGUACCCCCGGC





3103
GGAGGCGGUACCCCCGGCA





3104
GAGGCGGUACCCCCGGCAC





3105
AGGCGGUACCCCCGGCACC





3106
GGCGGUACCCCCGGCACCA





3107
GCGGUACCCCCGGCACCAU





3108
CGGUACCCCCGGCACCAUG





3109
GGUACCCCCGGCACCAUGA





3110
GUACCCCCGGCACCAUGAA





3111
UACCCCCGGCACCAUGAAU





3112
ACCCCCGGCACCAUGAAUG





3113
CCCCCGGCACCAUGAAUGG





3114
CCCCGGCACCAUGAAUGGC





3115
CCCGGCACCAUGAAUGGCA





3116
CCGGCACCAUGAAUGGCAU





3117
CGGCACCAUGAAUGGCAUC





3118
GGCACCAUGAAUGGCAUCC





3119
GCACCAUGAAUGGCAUCCA





3120
CACCAUGAAUGGCAUCCAU





3121
ACCAUGAAUGGCAUCCAUU





3122
CCAUGAAUGGCAUCCAUUG





3123
CAUGAAUGGCAUCCAUUGG





3124
AUGAAUGGCAUCCAUUGGG





3125
UGAAUGGCAUCCAUUGGGG





3126
GAAUGGCAUCCAUUGGGGC





3127
AAUGGCAUCCAUUGGGGCA





3128
AUGGCAUCCAUUGGGGCAG





3129
UGGCAUCCAUUGGGGCAGU





3130
GGCAUCCAUUGGGGCAGUG





3131
GCAUCCAUUGGGGCAGUGG





3132
CAUCCAUUGGGGCAGUGGG





3133
AUCCAUUGGGGCAGUGGGA





3134
UCCAUUGGGGCAGUGGGAC





3135
CCAUUGGGGCAGUGGGACA





3136
CAUUGGGGCAGUGGGACAC





3137
AUUGGGGCAGUGGGACACU





3138
UUGGGGCAGUGGGACACUG





3139
UGGGGCAGUGGGACACUGC





3140
GGGGCAGUGGGACACUGCA





3141
GGGCAGUGGGACACUGCAG





3142
GGCAGUGGGACACUGCAGG





3143
GCAGUGGGACACUGCAGGA





3144
CAGUGGGACACUGCAGGAC





3145
AGUGGGACACUGCAGGACC





3146
GUGGGACACUGCAGGACCA





3147
UGGGACACUGCAGGACCAG





3148
GGGACACUGCAGGACCAGA





3149
GGACACUGCAGGACCAGAG





3150
GACACUGCAGGACCAGAGG





3151
ACACUGCAGGACCAGAGGU





3152
CACUGCAGGACCAGAGGUG





3153
ACUGCAGGACCAGAGGUGA





3154
CUGCAGGACCAGAGGUGAG





3155
UGCAGGACCAGAGGUGAGG





3156
GCAGGACCAGAGGUGAGGA





3157
CAGGACCAGAGGUGAGGAU





3158
AGGACCAGAGGUGAGGAUG





3159
GGACCAGAGGUGAGGAUGA





3160
GACCAGAGGUGAGGAUGAA





3161
ACCAGAGGUGAGGAUGAAC





3162
CCAGAGGUGAGGAUGAACC





3163
CAGAGGUGAGGAUGAACCA





3164
AGAGGUGAGGAUGAACCAA





3165
GAGGUGAGGAUGAACCAAG





3166
AGGUGAGGAUGAACCAAGG





3167
GGUGAGGAUGAACCAAGGA





3168
GUGAGGAUGAACCAAGGAU





3169
UGAGGAUGAACCAAGGAUG





3170
GAGGAUGAACCAAGGAUGG





3171
AGGAUGAACCAAGGAUGGG





3172
GGAUGAACCAAGGAUGGGG





3173
GAUGAACCAAGGAUGGGGG





3174
AUGAACCAAGGAUGGGGGC





3175
UGAACCAAGGAUGGGGGCA





3176
GAACCAAGGAUGGGGGCAU





3177
AACCAAGGAUGGGGGCAUG





3178
ACCAAGGAUGGGGGCAUGG





3179
CCAAGGAUGGGGGCAUGGG





3180
CAAGGAUGGGGGCAUGGGG





3181
AAGGAUGGGGGCAUGGGGC





3182
AGGAUGGGGGCAUGGGGCC





3183
GGAUGGGGGCAUGGGGCCU





3184
GAUGGGGGCAUGGGGCCUU





3185
AUGGGGGCAUGGGGCCUUG





3186
UGGGGGCAUGGGGCCUUGG





3187
GGGGGCAUGGGGCCUUGGG





3188
GGGGCAUGGGGCCUUGGGA





3189
GGGCAUGGGGCCUUGGGAC





3190
GGCAUGGGGCCUUGGGACA





3191
GCAUGGGGCCUUGGGACAC





3192
CAUGGGGCCUUGGGACACU





3193
AUGGGGCCUUGGGACACUU





3194
UGGGGCCUUGGGACACUUG





3195
GGGGCCUUGGGACACUUGC





3196
GGGCCUUGGGACACUUGCC





3197
GGCCUUGGGACACUUGCCU





3198
GCCUUGGGACACUUGCCUU





3199
CCUUGGGACACUUGCCUUG





3200
CUUGGGACACUUGCCUUGU





3201
UUGGGACACUUGCCUUGUG





3202
UGGGACACUUGCCUUGUGC





3203
GGGACACUUGCCUUGUGCC





3204
GGACACUUGCCUUGUGCCC





3205
GACACUUGCCUUGUGCCCA





3206
ACACUUGCCUUGUGCCCAG





3207
CACUUGCCUUGUGCCCAGC





3208
ACUUGCCUUGUGCCCAGCC





3209
CUUGCCUUGUGCCCAGCCA





3210
UUGCCUUGUGCCCAGCCAG





3211
UGCCUUGUGCCCAGCCAGC





3212
GCCUUGUGCCCAGCCAGCC





3213
CCUUGUGCCCAGCCAGCCA





3214
CUUGUGCCCAGCCAGCCAC





3215
UUGUGCCCAGCCAGCCACG





3216
UGUGCCCAGCCAGCCACGU





3217
GUGCCCAGCCAGCCACGUG





3218
UGCCCAGCCAGCCACGUGG





3219
GCCCAGCCAGCCACGUGGA





3220
CCCAGCCAGCCACGUGGAC





3221
CCAGCCAGCCACGUGGACU





3222
CAGCCAGCCACGUGGACUG





3223
AGCCAGCCACGUGGACUGG





3224
GCCAGCCACGUGGACUGGA





3225
CCAGCCACGUGGACUGGAU





3226
CAGCCACGUGGACUGGAUU





3227
AGCCACGUGGACUGGAUUU





3228
GCCACGUGGACUGGAUUUC





3229
CCACGUGGACUGGAUUUCU





3230
CACGUGGACUGGAUUUCUG





3231
ACGUGGACUGGAUUUCUGC





3232
CGUGGACUGGAUUUCUGCU





3233
GUGGACUGGAUUUCUGCUC





3234
UGGACUGGAUUUCUGCUCU





3235
GGACUGGAUUUCUGCUCUU





3236
GACUGGAUUUCUGCUCUUC





3237
ACUGGAUUUCUGCUCUUCC





3238
CUGGAUUUCUGCUCUUCCA





3239
UGGAUUUCUGCUCUUCCAG





3240
GGAUUUCUGCUCUUCCAGA





3241
GAUUUCUGCUCUUCCAGAC





3242
AUUUCUGCUCUUCCAGACC





3243
UUUCUGCUCUUCCAGACCG





3244
UUCUGCUCUUCCAGACCGG





3245
UCUGCUCUUCCAGACCGGG





3246
CUGCUCUUCCAGACCGGGG





3247
UGCUCUUCCAGACCGGGGA





3248
GCUCUUCCAGACCGGGGAA





3249
CUCUUCCAGACCGGGGAAC





3250
UCUUCCAGACCGGGGAACU





3251
CUUCCAGACCGGGGAACUG





3252
UUCCAGACCGGGGAACUGG





3253
UCCAGACCGGGGAACUGGA





3254
CCAGACCGGGGAACUGGAC





3255
CAGACCGGGGAACUGGACU





3256
AGACCGGGGAACUGGACUC





3257
GACCGGGGAACUGGACUCA





3258
ACCGGGGAACUGGACUCAC





3259
CCGGGGAACUGGACUCACA





3260
CGGGGAACUGGACUCACAU





3261
GGGGAACUGGACUCACAUA





3262
GGGAACUGGACUCACAUAA





3263
GGAACUGGACUCACAUAAA





3264
GAACUGGACUCACAUAAAG





3265
AACUGGACUCACAUAAAGG





3266
ACUGGACUCACAUAAAGGC





3267
CUGGACUCACAUAAAGGCA





3268
UGGACUCACAUAAAGGCAA





3269
GGACUCACAUAAAGGCAAU





3270
GACUCACAUAAAGGCAAUG





3271
ACUCACAUAAAGGCAAUGU





3272
CUCACAUAAAGGCAAUGUC





3273
UCACAUAAAGGCAAUGUCC





3274
CACAUAAAGGCAAUGUCCU





3275
ACAUAAAGGCAAUGUCCUC





3276
CAUAAAGGCAAUGUCCUCU





3277
AUAAAGGCAAUGUCCUCUU





3278
UAAAGGCAAUGUCCUCUUU





3279
AAAGGCAAUGUCCUCUUUC





3280
AAGGCAAUGUCCUCUUUCU





3281
AGGCAAUGUCCUCUUUCUC





3282
GGCAAUGUCCUCUUUCUCU





3283
GCAAUGUCCUCUUUCUCUU





3284
CAAUGUCCUCUUUCUCUUC





3285
AAUGUCCUCUUUCUCUUCC





3286
AUGUCCUCUUUCUCUUCCC





3287
UGUCCUCUUUCUCUUCCCC





3288
GUCCUCUUUCUCUUCCCCC





3289
UCCUCUUUCUCUUCCCCCC





3290
CCUCUUUCUCUUCCCCCCA





3291
CUCUUUCUCUUCCCCCCAA





3292
UCUUUCUCUUCCCCCCAAC





3293
CUUUCUCUUCCCCCCAACC





3294
UUUCUCUUCCCCCCAACCC





3295
UUCUCUUCCCCCCAACCCU





3296
UCUCUUCCCCCCAACCCUU





3297
CUCUUCCCCCCAACCCUUU





3298
UCUUCCCCCCAACCCUUUA





3299
CUUCCCCCCAACCCUUUAU





3300
UUCCCCCCAACCCUUUAUU





3301
UCCCCCCAACCCUUUAUUU





3302
CCCCCCAACCCUUUAUUUU





3303
CCCCCAACCCUUUAUUUUG





3304
CCCCAACCCUUUAUUUUGU





3305
CCCAACCCUUUAUUUUGUG





3306
CCAACCCUUUAUUUUGUGU





3307
CAACCCUUUAUUUUGUGUU





3308
AACCCUUUAUUUUGUGUUU





3309
ACCCUUUAUUUUGUGUUUU





3310
CCCUUUAUUUUGUGUUUUA





3311
CCUUUAUUUUGUGUUUUAA





3312
CUUUAUUUUGUGUUUUAAG





3313
UUUAUUUUGUGUUUUAAGC





3314
UUAUUUUGUGUUUUAAGCU





3315
UAUUUUGUGUUUUAAGCUG





3316
AUUUUGUGUUUUAAGCUGU





3317
UUUUGUGUUUUAAGCUGUA





3318
UUUGUGUUUUAAGCUGUAU





3319
UUGUGUUUUAAGCUGUAUG





3320
UGUGUUUUAAGCUGUAUGA





3321
GUGUUUUAAGCUGUAUGAC





3322
UGUUUUAAGCUGUAUGACU





3323
GUUUUAAGCUGUAUGACUU





3324
UUUUAAGCUGUAUGACUUU





3325
UUUAAGCUGUAUGACUUUA





3326
UUAAGCUGUAUGACUUUAU





3327
UAAGCUGUAUGACUUUAUC





3328
AAGCUGUAUGACUUUAUCA





3329
AGCUGUAUGACUUUAUCAC





3330
GCUGUAUGACUUUAUCACU





3331
CUGUAUGACUUUAUCACUG





3332
UGUAUGACUUUAUCACUGA





3333
GUAUGACUUUAUCACUGAG





3334
UAUGACUUUAUCACUGAGA





3335
AUGACUUUAUCACUGAGAA





3336
UGACUUUAUCACUGAGAAU





3337
GACUUUAUCACUGAGAAUA





3338
ACUUUAUCACUGAGAAUAA





3339
CUUUAUCACUGAGAAUAAU





3340
UUUAUCACUGAGAAUAAUA





3341
UUAUCACUGAGAAUAAUAC





3342
UAUCACUGAGAAUAAUACA





3343
AUCACUGAGAAUAAUACAU





3344
UCACUGAGAAUAAUACAUG





3345
CACUGAGAAUAAUACAUGU





3346
ACUGAGAAUAAUACAUGUU





3347
CUGAGAAUAAUACAUGUUA





3348
UGAGAAUAAUACAUGUUAA





3349
GAGAAUAAUACAUGUUAAA





3350
AGAAUAAUACAUGUUAAAC





3351
GAAUAAUACAUGUUAAACG





3352
AAUAAUACAUGUUAAACGU





3353
AUAAUACAUGUUAAACGUU





3354
UAAUACAUGUUAAACGUUU





3355
AAUACAUGUUAAACGUUUG





3356
AUACAUGUUAAACGUUUGU





3357
UACAUGUUAAACGUUUGUG





3358
ACAUGUUAAACGUUUGUGG





3359
CAUGUUAAACGUUUGUGGU





3360
AUGUUAAACGUUUGUGGUA





3361
UGUUAAACGUUUGUGGUAA





3362
GUUAAACGUUUGUGGUAAG





3363
UUAAACGUUUGUGGUAAGA





3364
UAAACGUUUGUGGUAAGAG





3365
AAACGUUUGUGGUAAGAGG





3366
AACGUUUGUGGUAAGAGGU





3367
ACGUUUGUGGUAAGAGGUC





3368
CGUUUGUGGUAAGAGGUCA





3369
GUUUGUGGUAAGAGGUCAG





3370
UUUGUGGUAAGAGGUCAGU





3371
UUGUGGUAAGAGGUCAGUG





3372
UGUGGUAAGAGGUCAGUGG





3373
GUGGUAAGAGGUCAGUGGU





3374
UGGUAAGAGGUCAGUGGUA





3375
GGUAAGAGGUCAGUGGUAU





3376
GUAAGAGGUCAGUGGUAUC





3377
UAAGAGGUCAGUGGUAUCU





3378
AAGAGGUCAGUGGUAUCUG





3379
AGAGGUCAGUGGUAUCUGC





3380
GAGGUCAGUGGUAUCUGCC





3381
AGGUCAGUGGUAUCUGCCC





3382
GGUCAGUGGUAUCUGCCCU





3383
GUCAGUGGUAUCUGCCCUG





3384
UCAGUGGUAUCUGCCCUGA





3385
CAGUGGUAUCUGCCCUGAA





3386
AGUGGUAUCUGCCCUGAAU





3387
GUGGUAUCUGCCCUGAAUC





3388
UGGUAUCUGCCCUGAAUCU





3389
GGUAUCUGCCCUGAAUCUG





3390
GUAUCUGCCCUGAAUCUGC





3391
UAUCUGCCCUGAAUCUGCU





3392
AUCUGCCCUGAAUCUGCUU





3393
UCUGCCCUGAAUCUGCUUC





3394
CUGCCCUGAAUCUGCUUCA





3395
UGCCCUGAAUCUGCUUCAA





3396
GCCCUGAAUCUGCUUCAAA





3397
CCCUGAAUCUGCUUCAAAG





3398
CCUGAAUCUGCUUCAAAGA





3399
CUGAAUCUGCUUCAAAGAG





3400
UGAAUCUGCUUCAAAGAGU





3401
GAAUCUGCUUCAAAGAGUU





3402
AAUCUGCUUCAAAGAGUUA





3403
AUCUGCUUCAAAGAGUUAU





3404
UCUGCUUCAAAGAGUUAUU





3405
CUGCUUCAAAGAGUUAUUU





3406
UGCUUCAAAGAGUUAUUUC





3407
GCUUCAAAGAGUUAUUUCA





3408
CUUCAAAGAGUUAUUUCAA





3409
UUCAAAGAGUUAUUUCAAA





3410
UCAAAGAGUUAUUUCAAAU





3411
CAAAGAGUUAUUUCAAAUU





3412
AAAGAGUUAUUUCAAAUUA





3413
AAGAGUUAUUUCAAAUUAA





3414
AGAGUUAUUUCAAAUUAAA





3415
GAGUUAUUUCAAAUUAAAA





3416
AGUUAUUUCAAAUUAAAAG





3417
GUUAUUUCAAAUUAAAAGC





3418
UUAUUUCAAAUUAAAAGCA





3419
UAUUUCAAAUUAAAAGCAA





3420
AUUUCAAAUUAAAAGCAAA





3421
UUUCAAAUUAAAAGCAAAA





3422
UUCAAAUUAAAAGCAAAAC





3423
UCAAAUUAAAAGCAAAACA





3424
CAAAUUAAAAGCAAAACAA





3425
AAAUUAAAAGCAAAACAAA





3426
AAUUAAAAGCAAAACAAAA





3427
AUUAAAAGCAAAACAAAAC





3428
UUAAAAGCAAAACAAAACA





3429
UAAAAGCAAAACAAAACAA





3430
AAAAGCAAAACAAAACAAA









The double-stranded regions should be at least 19 contiguous nucleotides, for example about 19 to 23 nucleotides, or may be longer, for example 30 or 50 nucleotides, or 100 nucleotides or more. The full-length sequence corresponding to the entire gene transcript may be used. Preferably, they are about 19 to about 100 nucleotides in length, more preferably about 19 to about 50 nucleotides in length, and even more preferably about 19 to about 23 nucleotides in length.


The degree of identity of a double-stranded region of a nucleic acid molecule to the targeted transcript should be at least 90% and more preferably 95-100%. The % identity of a nucleic acid molecule is determined by GAP (Needleman and Wunsch, 1970) analysis (GCG program) with a gap creation penalty=5, and a gap extension penalty=0.3. Preferably, the two sequences are aligned over their entire length.


The nucleic acid molecule may of course comprise sequences unrelated to the target which may function to stabilize the molecule.


The term “short interfering RNA” or “siRNA” as used herein refers to a nucleic acid molecule which comprises ribonucleotides capable of inhibiting or down regulating gene expression, for example by mediating RNAi in a sequence-specific manner, wherein the double stranded portion is less than 50 nucleotides in length, preferably about 19 to about 23 nucleotides in length. For example, the siRNA can be a nucleic acid molecule comprising self-complementary sense and antisense regions, wherein the antisense region comprises a nucleotide sequence that is complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense region having nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. The siRNA can be assembled from two separate oligonucleotides, where one strand is the sense strand and the other is the antisense strand, wherein the antisense and sense strands are self-complementary.


As used herein, the term siRNA is equivalent to other terms used to describe nucleic acid molecules that are capable of mediating sequence specific RNAi, for example micro-RNA (miRNA), short hairpin RNA (shRNA), short interfering oligonucleotide, short interfering nucleic acid (siNA), short interfering modified oligonucleotide, chemically-modified siRNA, and others. In addition, as used herein, the term RNAi is equivalent to other terms used to describe sequence specific RNA interference, such as post transcriptional gene silencing, translational inhibition, or epigenetics. For example, siRNA molecules of the invention can be used to epigenetically silence genes at both the post-transcriptional level or the pre-transcriptional level. In a non-limiting example, epigenetic regulation of gene expression by siRNA molecules of the invention can result from siRNA mediated modification of chromatin structure to alter gene expression.


Preferred siRNA molecules comprise a nucleotide sequence that is identical to about 19 to 23 contiguous nucleotides of the target mRNA. In an embodiment, the target mRNA sequence commences with the dinucleotide AA, comprises a GC-content of about 30-70% (preferably, 30-60%, more preferably 40-60% and more preferably about 45%-55%), and does not have a high percentage identity to any nucleotide sequence other than the target in the genome of the avian (preferably chickens) in which it is to be introduced, e.g., as determined by standard BLAST search.


By “shRNA” or “short-hairpin RNA” is meant an siRNA molecule where less than about 50 nucleotides, preferably about 19 to about 23 nucleotides, is base paired with a complementary sequence located on the same RNA molecule, and where said sequence and complementary sequence are separated by an unpaired region of at least about 4 to 15 nucleotides which forms a single-stranded loop above the stem structure created by the two regions of base complementarity. Examples of sequences of a single-stranded loops are 5′ UUCAAGAGA 3′ and 5′ UUUGUGUAG 3′.


Included shRNAs are dual or bi-finger and multi-finger hairpin dsRNAs, in which the RNA molecule comprises two or more of such stem-loop structures separated by single-stranded spacer regions.


siRNAs can be generated in vitro by using a recombinant enzyme, such as T7 RNA polymerase, and DNA oligonucleotide templates, or can be prepared in vivo, for example, in cultured cells. In a preferred embodiment, the nucleic acid molecule is produced synthetically.


Strategies have been described for producing a hairpin siRNA from vectors containing, for example, a RNA polymerase III promoter. Various vectors have been constructed for generating hairpin siRNAs in host cells using either an H1-RNA or an snU6 RNA promoter (see SEQ ID NO's 7 to 9). A RNA molecule as described above (e.g., a first portion, a linking sequence, and a second portion) can be operably linked to such a promoter. When transcribed by RNA polymerase III, the first and second portions form a duplexed stem of a hairpin and the linking sequence forms a loop. The pSuper vector (OligoEngines Ltd., Seattle, Wash.) can also be used to generate siRNA.


Modifications or analogs of nucleotides can be introduced to improve the properties of the nucleic acid molecules of the invention. Improved properties include increased nuclease resistance and/or increased ability to permeate cell membranes. Accordingly, the terms “nucleic acid molecule” and “double-stranded RNA molecule” includes synthetically modified bases such as, but not limited to, inosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl-, 2-propyl- and other alkyl-adenines, 5-halo uracil, 5-halo cytosine, 6-aza cytosine and 6-aza thymine, pseudo uracil, 4-thiuracil, 8-halo adenine, 8-aminoadenine, 8-thiol adenine, 8-thiolalkyl adenines, 8-hydroxyl adenine and other 8-substituted adenines, 8-halo guanines, 8-amino guanine, 8-thiol guanine, 8-thioalkyl guanines, 8-hydroxyl guanine and other substituted guanines, other aza and deaza adenines, other aza and deaza guanines, 5-trifluoromethyl uracil and 5-trifluoro cytosine.


Vectors and Host Cells

The present invention also provides a vector encoding a nucleic acid molecule comprising a double-stranded region, or single strand thereof, of the present invention. Preferably, the vector is an expression vector capable of expressing the open reading frame(s) encoding a dsRNA in a host cell and/or cell free system. The host cell can be any cell type such as, not limited to, bacterial, fungal, plant or animal cells, preferably an avian cell.


Typically, a vector of the invention comprises a promoter operably linked to an open reading frame encoding a nucleic acid molecule of the invention, or a strand thereof.


As used herein, the term “promoter” refers to a nucleic acid sequence which is able to direct transcription of an operably linked nucleic acid molecule and includes, for example, RNA polymerase II and RNA polymerase III promoters. Also included in this definition are those transcriptional regulatory elements (e.g., enhancers) that are sufficient to render promoter-dependent gene expression controllable in a cell type-specific, tissue-specific, or temporal-specific manner, or that are inducible by external agents or signals.


“Operably linked” as used herein refers to a functional relationship between two or more nucleic acid (e.g., DNA) segments. Typically, it refers to the functional relationship of a transcriptional regulatory element to a transcribed sequence. For example, a promoter is operably linked to a coding sequence, such as an open reading frame encoding a double-stranded RNA molecule defined herein, if it stimulates or modulates the transcription of the coding sequence in an appropriate cell. Generally, promoter transcriptional regulatory elements that are operably linked to a transcribed sequence are physically contiguous to the transcribed sequence, i.e., they are cis-acting. However, some transcriptional regulatory elements, such as enhancers, need not be physically contiguous or located in close proximity to the coding sequences whose transcription they enhance.


By “RNA polymerase III promoter” or “RNA pol III promoter” or “polymerase III promoter” or “pol III promoter” is meant any invertebrate, vertebrate, or mammalian promoter, e.g., chicken, human, murine, porcine, bovine, primate, simian, etc. that, in its native context in a cell, associates or interacts with RNA polymerase III to transcribe its operably linked gene, or any variant thereof, natural or engineered, that will interact in a selected host cell with an RNA polymerase III to transcribe an operably linked nucleic acid sequence. By U6 promoter (e.g., chicken U6, human U6, murine U6), H1 promoter, or 7SK promoter is meant any invertebrate, vertebrate, or mammalian promoter or polymorphic variant or mutant found in nature to interact with RNA polymerase III to transcribe its cognate RNA product, i.e., U6 RNA, H1 RNA, or 7SK RNA, respectively. Examples of suitable promoters include cU6-1 (SEQ ID NO:7), cU6-3 (SEQ ID NO:8), cU6-4 (SEQ ID NO:9) and c7SK (SEQ ID NO:10).


When E. coli is used as a host cell, there is no limitation other than that the vector should have an “ori” to amplify and mass-produce the vector in E. coli (e.g., JM109. DH5α, HB101, or XL1Blue), and a marker gene for selecting the transformed E. coli (e.g., a drug-resistance gene selected by a drug such as ampicillin, tetracycline, kanamycin, or chloramphenicol). For example, M13-series vectors, pUC-series vectors, pBR322, pBluescript, pCR-Script, and such can be used. pGEM-T, pDIRECT, pT7, and so on can also be used for subcloning and excision of the gene encoding the dsRNA as well as the vectors described above.


With regard to expression vectors for use in E. coli, such vectors include JM109, DH5α, HB101, or XL1 Blue, the vector should have a promoter such as lacZ promoter, araB promoter, or T7 promoter that can efficiently promote the expression of the desired gene in E. coli. Other examples of the vectors are “QIAexpress system” (Qiagen), pEGFP, and pET (for this vector, BL21, a strain expressing T7 RNA polymerase, is preferably used as the host).


In addition to the vectors for E. coli, for example, the vector may be a mammal-derived expression vector (e.g., pcDNA3 (Invitrogen), pEGF-BOS, pEF, and pCDM8), an insect cell-derived expression vector (e.g., “Bac-to-BAC baculovairus expression system” (GibcoBRL) and pBacPAK8), a plant-derived expression vector (e.g., pMH1 and pMH2), an animal virus-derived expression vector (e.g., pHSV, pMV, and pAdexLcw), a retrovirus-derived expression vector (e.g., pZIPneo), a yeast-derived expression vector (e.g., “Pichia Expression Kit” (Invitrogen), pNV11, and SP-Q01), or a Bacillus subtilis-derived expression vector (e.g., pPL608 and pKTH50).


In order to express nucleic acid molecules in animal cells, such as CHO, COS, Vero and NIH3T3 cells, the vector should have a promoter necessary for expression in such cells, e.g., SV40 promoter, MMLV-LTR promoter, EF1α promoter, CMV promoter, etc., and more preferably it has a marker gene for selecting transformants (for example, a drug resistance gene selected by a drug (e.g., neomycin, G418, etc.). Examples of vectors with these characteristics include pMAM, pDR2, pBK-RSV, pBK-CMV, pOPRSV and pOPI3.


Nucleic acid molecules comprising a double-stranded region of the present invention can be expressed in animals such as avians by, for example, inserting an open reading frame(s) encoding the nucleic acid into an appropriate vector and introducing the vector by the retrovirus method, liposome method, cationic liposome method, adenovirus method, and so on. The vectors used include, but are not limited to, adenoviral vectors (e.g., pAdexlcw) and retroviral vectors (e.g., pZIPneo). General techniques for gene manipulation, such as insertion of nucleic acids of the invention into a vector, can be performed according to conventional methods.


The present invention also provides a host cell into which an exogenous nucleic acid molecule, typically in a vector of the present invention, has been introduced. The host cell of this invention can be used as, for example, a production system for producing or expressing the nucleic acid molecule. For in vitro production, eukaryotic cells or prokaryotic cells can be used.


Useful eukaryotic host cells may be animal, plant, or fungi cells. As animal cells, mammalian cells such as CHO, COS, 3T3, myeloma, baby hamster kidney (BHK), HeLa, or Vero cells MDCK cells, DF1 cells, amphibian cells such as Xenopus oocytes, or insect cells such as Sf9, Sf21, or Tn5 cells can be used. CHO cells lacking DHFR gene (dhfr-CHO) or CHO K-1 may also be used. The vector can be introduced into the host cell by, for example, the calcium phosphate method, the DEAE-dextran method, cationic liposome DOTAP (Boehringer Mannheim) method, electroporation, lipofection, etc.


Useful prokaryotic cells include bacterial cells, such as E. coli, for example, JM109, DH5α, and HB101, or Bacillus subtilis.


Culture medium such as DMEM, MEM, RPMI-1640, or IMDM may be used for animal cells. The culture medium can be used with or without serum supplement such as fetal calf serum (FCS). The pH of the culture medium is preferably between about 6 and 8. Cells are typically cultured at about 30 to 40° C. for about 15 to 200 hr, and the culture medium may be replaced, aerated, or stirred if necessary.


Compositions

The present invention also provides compositions comprising a nucleic acid molecule comprising a double-stranded region that can be administered to an avian egg. A composition comprising a nucleic acid molecule comprising a double-stranded region may contain a pharmaceutically acceptable carrier to render the composition suitable for administration.


Suitable pharmaceutical carriers, excipients and/or diluents include, but are not limited to, lactose, sucrose, starch powder, talc powder, cellulose esters of alkonoic acids, magnesium stearate, magnesium oxide, crystalline cellulose, methyl cellulose, carboxymethyl cellulose, gelatin, glycerin, sodium alginate, antibacterial agents, antifungal agents, gum arabic, acacia gum, sodium and calcium salts of phosphoric and sulfuric acids, polyvinylpyrrolidone and/or polyvinyl alcohol, saline, and water. In an embodiment, the carrier, excipient and/or diluent is phosphate buffered saline or water.


In an embodiment, the composition may also comprise a transfection promoting agent. Transfection promoting agents used to facilitate the uptake of nucleic acids into a living cell are well known within the art. Reagents enhancing transfection include chemical families of the types; polycations, dendrimers, DEAE Dextran, block copolymers and cationic lipids. Preferably, the transfection-promoting agent is a lipid-containing compound (or formulation), providing a positively charged hydrophilic region and a fatty acyl hydrophobic region enabling self-assembly in aqueous solution into vesicles generally known as micelles or liposomes, as well as lipopolyamines.


In another embodiment, the composition comprises a polymeric biomaterial such as chitosan.


It is understood that any conventional media or agent may be used so long as it is not incompatible with the compositions or methods of the invention.


Administration

Administration of a nucleic acid molecule comprising a double-stranded region (including a composition comprising a nucleic acid molecule comprising a double-stranded region) is conveniently achieved by injection into the egg, and generally injection into the chorion allantoic fluid. Notwithstanding that the air sac is the preferred route of in ovo administration, other regions such as the yolk sac, air sac or amnionic cavity (amnion) may also be inoculated by injection. The hatchability rate might decrease slightly when the air sac is not the target for the administration although not necessarily at commercially unacceptable levels. The mechanism of injection is not critical to the practice of the present invention, although it is preferred that the needle does not cause undue damage to the egg or to the tissues and organs of the developing embryo or the extra-embryonic membranes surrounding the embryo.


Preferably, the nucleic acid molecule is administered within four days of the egg having been laid.


Generally, a hypodermic syringe fitted with an approximately 22 gauge needle is suitable. The method of the present invention is particularly well adapted for use with an automated injection system, such as those described in U.S. Pat. No. 4,903,635, U.S. Pat. No. 5,056,464, U.S. Pat. No. 5,136,979 and US 20060075973.


The nucleic acid molecule is administered in an effective amount sufficient to modify sex in at least some of the eggs which have been administered. The modification can be detected by comparing a suitable number of samples subjected to the method of the invention to a similar number that have not. Statistically significant variation in the sex of the birds between the two groups will be indicative that an effective amount has been administered. Other means of determining an effective amount for sex are well within the capacity of those skilled in the art.


Preferably, about 1 ng to 100 μg, more preferably about 100 ng to 1 μg, of nucleic acid is administered to the egg. Furthermore, it is preferred that the nucleic acid to be administered is in a volume of about 1 μl to 1 ml, more preferably about 10 μl to 500 μl.


EXAMPLES
Example 1
Identification of shRNA Molecules for Down-Regulating DMRT1 Protein Production in Chickens

Selection of shRNA Sequences Targeting DMRT1


The present inventors identified 51 predicted shRNA sequences to target chicken Dmrt1 (Table 4).


There are several algorithms available to select potential siRNA sequences for specific target genes. Taxman et al. (2006) have specifically designed an algorithm to predict effective shRNA molecules and the present inventors have made their own modification to the algorithm to improve shRNA prediction. There are four criteria for shRNA selection using the Taxman algorithm. Three of the criteria are scored for out of a maximum number of 4 points. These criteria are: 1) C or G on the 5′ end of the sequence=1 point, A or T on 5′ end=−1 point; 2) A or T on the 3′ end=1 point, C or G on the 3′ end=−1 point; 3) 5 or more A or T in the seven 3′ bases=2 points, 4 A or T in the seven 3′ bases=1 point. shRNA sequences with the highest scores are preferred. The fourth criteria is based on a calculation for the free-energy of the 6 central bases of the shRNA sequence (bases 6-11 of the sense strand hybridised to bases 9-14 of the antisense strand). shRNAs with a central duplex ΔG>−12.9 kcal/mol are preferred.


The shRNA designer website uses this algorithm to provide a score for each shRNA target. Based on the algorithm and their calculated ΔG value, the present inventors chose 4 of the shRNA target finder shRNA sequences as potentially effective shRNAs to test for their ability to knockdown Dmrt1 gene expression. The selected sequences are shown in their 5′-3′ sequence in Table 5. These 4 sequences were used to construct ddRNAi plasmids for the expression of the 6 shRNAs.


Construction of ddRNAi Plasmids for Expression of Selected shRNAs


To construct the Dmrt1 shRNA expression constructs, two oligonucleotides complementary to each other were designed to contain the sense, loop, antisense and termination signal, followed by a spacer sequence (GGAA) and a BamHI restriction site for screening. In addition, the “bottom oligo (B)” or reverse oligonucleotide contained a SalI overhang at the 5′ end for insertion into the expression vector containing the chicken polymerase III promoter cU6-4 (DQ531570). Table 6 lists the shRNA targets to their corresponding oligonucleotides. The complementary oligonucleotides for each target shRNA, were annealed together and ligated into the PmeI-SalI digested pU6-4 vector. Full length clones were positive if linearised by BamHI digestion. All shRNA expression vectors were sequence confirmed. The four constructs were referred to as 105sh, 240sh, 465sh and 591sh as shown in FIG. 1. The non-silencing control shRNA construct (NSsh) was designed in the same way. However, the sequence used was that of an irrelevant target (ie. NP gene of influenza).


Each ddRNAi plasmid was constructed so that the start of each shRNA sequence was at the +1 position of the native U6 snRNA transcripts. All final shRNA expression vectors consisted of either one of the full length chicken U6 promoters, a shRNA sense sequence, a loop sequence, a shRNA antisense sequence, a termination sequence and a BamHI site. The loop sequence used in all shRNAs was 5′ UUCAAGAGA 3′.









TABLE 4







Algorithm selection of shRNA sequences targeting


Dmrt1.










Target Sequence
Start Position
SEQ ID NO
Score













GGCACAAGCGGTTCTGCAT
79
3435
3





GACTGCCAGTGCAAGAAGT
105
3436
3





CTGAGCCAGTTGTCAAGAA
238
3437
3





GAGCCAGTTGTCAAGAAGA
240
3438
3





GACGGATGCTCATTCAGGA
355
3439
3





GCACGTCTGATTTGGTTGT
409
3440
3





GTGGACTCCACCTACTACA
426
3441
3





CCAGCCATCCCTGTATCCT
455
3442
3





CCATCCCTGTATCCTTACT
459
3443
3





CATCCCTGTATCCTTACTA
460
3444
3





CCCTGTATCCTTACTATAA
463
3445
4





CTGTATCCTTACTATAACA
465
3446
4





CTTACTATAACAACCTGTA
472
3447
3





CTCCCAGTACCAAATGGCA
497
3448
3





GCCACTGAGTCTTCCTCAA
519
3449
3





CTGAGTCTTCCTCAAGTGA
523
3450
3





GAGTCTTCCTCAAGTGAGA
525
3451
3





CTCCCAGCAACATACATGT
591
3452
4





CCCAGCAACATACATGTCA
593
3453
3





CCAGCAACATACATGTCAA
594
3454
3





CAGATGAAGGGAATGGAGA
633
3455
3





CCACCTGCGTCACACAGAT
733
3456
3





CACCTGCGTCACACAGATA
734
3457
3





CCTGCGTCACACAGATACT
736
3458
3





CTCCTACTCAGAGTCGAAA
773
3459
3





CACTGTTGCCTTGTTCTGT
967
3460
3





GGTGCCGTGATGTGTTTGT
1189
3461
3





GTGCCGTGATGTGTTTGTA
1190
3462
4





GCCGTGATGTGTTTGTAGT
1192
3463
4





CCTCGTATCGCCAAATTAA
1239
3464
4





GCCTCGACTTAGATTGCAA
1283
3465
3





CCTCGACTTAGATTGCAAT
1284
3466
4





CTCGACTTAGATTGCAATA
1285
3467
4





CGACTTAGATTGCAATATA
1287
3468
4





GACTTAGATTGCAATATAA
1288
3469
4





GCGGCCAGCAAACAAGTCT
1307
3470
3





GGCCAGCAAACAAGTCTCA
1309
3471
3





GCCAGCAAACAAGTCTCAA
1310
3472
3





CCAGCAAACAAGTCTCAAA
1311
3473
3





GCGTTTCTGCGAGTGTTAT
1342
3474
4





GTGTCCTCTTCCTGTGTTA
1381
3475
3





GTCCTCTTCCTGTGTTACA
1383
3476
3





CCTCTTCCTGTGTTACAGA
1385
3477
3





CTCTTCCTGTGTTACAGAA
1386
3478
4





GAAGCCAACCTGAAATGAA
1402
3479
4





GCCAACCTGAAATGAAACT
1405
3480
4





CCAACCTGAAATGAAACTA
1406
3481
4





CCTGAAATGAAACTAGTCT
1410
3482
3





GTTGCAGCTGTACCTGAAA
1452
3483
3





GCAGCTGTACCTGAAATAA
1455
3484
4





CAGCTGTACCTGAAATAAA
1456
3485
4
















TABLE 5







Sequence of Dmrt1 shRNAs.











Target






Number
Position
Target Sequence
Score
ΔG





1
105 bp
GACTGCCAGTGCAAGAAGT
3
−17.1




(SEQ ID NO: 3436)





2
240 bp
GAGCCAGTTGTCAAGAAGA
3
−12.7




(SEQ ID NO: 3438)





3
465 bp
CTGTATCCTTACTATAACA
4
−11.9




(SEQ ID NO: 3446)





4
591 bp
CTCCCAGCAACATACATGT
4
−14.0




(SEQ ID NO: 3452)










Testing Selected shRNAs for Knockdown of Dmrt1 Gene Expression


A reporter gene expression assay was used to test shRNAs for silencing of Dmrt1. The reporter gene was a transcriptional gene fusion of Dmrt1 inserted downstream of the 3′ end of the Enhanced Green Fluorescent Protein (EGFP) gene in pEGFP-C (Clontech). The reporter plasmid was constructed as follows: cDNA of Dmrt1 was reverse transcribed from total RNA isolated from 4 day old embryo's and cloned into the multiple cloning site of pCMV-Script (Stratagene). The Dmrt1 insert was removed from the cloning vector as a NotI-EcoRI fragment and cloned downstream of the EGFP gene in pEGFP-C (Clontech). The resulting plasmid was named pEGFP-Dmrt1. This plasmid was transfected into chicken DF-1 cells and expression of the transcriptional gene fusion was confirmed by measuring EGFP fluorescence using flow cytometry as described below.


Dmrt1 gene silencing assays were conducted by co-transfecting DF-1 cells with the pEGFP-Dmrt1 reporter plasmid and each of the ddRNAi plasmids expressing the Dmrt1 specific and control shRNAs. The co-transfection experiments were performed as follows: DF-1 cells (ATCC CRL-12203, chicken fibroblast) were maintained in a humidified atmosphere containing 5% CO2 at 37° C. in Dulbecco's Modified Eagle's Medium (DMEM) containing 4.5 g/l glucose, 1.5 g/l sodium bicarbonate, 10% foetal calf serum (FCS), 2 mM L-glutamine supplemented with penicillin (100 Um') and streptomycin (100 μg/ml). DF1 cells were passaged as required using 0.25% (w/v) trypsin-ethylenediaminetetraacetic acid (EDTA).









TABLE 6





Sequence and details of primers used.



















Target






Number
Position
Target Sequence
Score
ΔG





1
105 bp
GACTGCCAGTGCAAGAAGT
3
−17.1




(SEQ ID NO: 3436)







DMRT1-105T-U6.4


GACTGCCAGTGCAAGAAGTTTCAAGAGAACTTCTTGCACTGGCAGTCTTT


TTGGAAGGATCC (SEQ ID NO: 3486)


DMRT1-105B-U6.4



TCGA
GGATCC
TTCCAAAAAGACTGCCAGTGCAAGAAGTTCTCTTGAAAC



TTCTTGCACTGGCAGTC (SEQ ID NO: 3487)














Target






Number
Position
Target Sequence
Score
ΔG





2
240 bp
GAGCCAGTTGTCAAGAAGA
3
−12.7




(SEQ ID NO: 3438)







DMRT1-240T-U6.4


GAGCCAGTTGTCAAGAAGATTCAAGAGATCTTCTTGACAACTGGCTCTTT


TTGGAAGGATCC (SEQ ID NO: 3488)


DMRT1-240B-U6.4



TCGA
GGATCC
TTCCAAAAAGAGCCAGTTGTCAAGAAGATCTCTTGAATCT



TCTTGACAACTGGCTC (SEQ ID NO: 3489)














Target






Number
Position
Target Sequence
Score
ΔG





3
465 bp
CTGTATCCTTACTATAACA
4
−11.9




(SEQ ID NO: 3446)







DMRT1-465T-U6.4


CTGTATCCTTACTATAACATTCAAGAGATGTTATAGTAAGGATACAGTTT


TTGGAAGGATCC (SEQ ID NO: 3490)


DMRT1-465B-U6.4



TCGA
GGATCC
TTCCAAAAACTGTATCCTTACTATAACATCTCTTGAATGT



TATAGTAAGGATACAG (SEQ ID NO: 3491)














Target






Number
Position
Target Sequence
Score
ΔG





4
591 bp
CTCCCAGCAACATACATGT
4
−14.0




(SEQ ID NO: 3452)







DMRT1-591T-U6.4


CTCCCAGCAACATACATGTTTCAAGAGAACATGTATGTTGCTGGGAGTTT


TTGGAAGGATCC (SEQ ID NO: 3492)


DMRT1-591B-U6.4



TCGA
GGATCC
TTCCAAAAACTCCCAGCAACATACATGTTCTCTTGAAACA



TGTATGTTGCTGGGAG (SEQ ID NO: 3493)





Bold = spacer sequence;


Italics = BamHI restriction site;


Underline = SalI overhang






Co-transfection of pEGFP-Dmrt1 and ddRNAi plasmids for EGFP-Dmrt1 fusion silencing assays was conducted in DF-1 cells grown to 80-90% confluence, in 24 well culture plates (Nunc) for flow cytometry analysis. Cells were transfected with a total of 1 μg of plasmid DNA, per well, using Lipofectamine™2000 transfection reagent (Invitrogen). EGFP expression was analysed in transfected DF-1 cells at 60 hours post-transfection using flow cytometry analysis of transfections performed in triplicate. Cells were trypsinised using 100 μl of 0.25% trypsin-EDTA, pelleted at 2000 rpm for 5 minutes, washed once in 1 ml of cold phosphate buffered saline-A (PBSA) (Oxoid), twice in 1 ml of FACS-wash solution (PBSA+1% FCS) and resuspended in 250 μl of FACS-wash solution. Flow cytometry sampling was performed using a FACScalibur (Becton Dickinson) fluorescence activated cell sorter. Data acquisition and calculation of mean fluorescence intensity (MFI) values for triplicate co-transfection samples, was performed using CELLQuest software (Becton Dickinson). The results of the gene silencing assay are shown in FIG. 1. Compared to the negative control irrelevant shRNA expressed from NSsh, the Dmrt1 specific shRNAs were observed to knockdown expression of the reporter gene to varying levels. Dmrt1 shRNA 240sh induced the greatest level of gene silencing of approximately 60%.


Example 2
In Ovo Modulation of DMRT1 Gene Expression in Chickens

An siRNA targeted to a conserved exon of the chicken DMRT1 gene was designed using the Ambion siRNA Target Finder tool (www.ambion.com). The chosen siRNA was designated DMRT1-343-siRNA (5′-GAGCCAGUUGUCAAGAAGAUU-3′) (SEQ ID NO:3431). The siRNA was synthesized and obtained from Qiagen.


For in ovo delivery, the siRNA was formulated with lipoefectamine 2000 (Invitrogen) according to the manufacturer's instructions. The now complexed siRNA was then delivered in ovo at a dose of either 100 μmol or 200 μmol. The siRNA was injected into embryonated eggs via an intravenous (I.V.) route or directly into the amnion at embryonic day 4.5 (E4.5). For both I.V. and amnion delivery, a small opening (1 cm×1 cm) was created at the top of the blunt end of the egg so as to avoid the membrane, veins and arteries, and 100 pmol or 200 pmol in a 4 μl volume was then injected directly into a vein or into the amnionic cavity using a micro-capillary pipette. Micro-capillaries of 1 mm diameter were used for injections, and their tips were pulled to a diameter of 40 microns with bevelled tip of 22.5°. After injection, the holes in the eggs were sealed with appropriate sized parafilm squares using a heated scalpel blade.


In total, 286 embryonated eggs (E4.5) were used in this experiment;


Group 1: 48 eggs were used as controls and were not injected with the DMRT1-343-siRNA formulation;


Group 2: 51 eggs were injected I.V. with 100 pmol of siRNA;


Group 3: 53 eggs were injected I.V. with 200 pmol of siRNA;


Group 4: 81 eggs were injected into the amnion with 100 pmol of siRNA and;


Group 5: 53 eggs were injected into the amnion with 200 pmol of siRNA.


All embryos were incubated until day E10. At E10, all embryos were assessed for viability and then removed from the egg. Control Group 1 had an embryo viability of 100%; Group 2 had a viability of 76%; Group 3 had a viability of 94%; Group 4 had a viability of 40% and; Group 5 had a viability of 75%. A single limb bud from each embryo was removed and used in a sex determination PCR test to determine if the embryos were of male or female genotype. Lower limb buds from each embryo were collected into 50 μl of PCR digestion buffer (50 mM KCl; 10 mM Tris-HCl, pH8.3; 0.1 mg/ml gelatine; 0.45% Nonidet P-40; 0.45% Tween-20; 0.2 mg/ml proteinase K; stock stored at −20° C.) at room temperature and digested at 55° C. for a minimum of 1 h, then at 95° C. for 10 min to release genomic DNA.


Sexing was carried out by PCR using the method of Clinton et al. (2001). The PCR mix consisted of 1 μl of digestion mix, 10×RedTaq reaction buffer (Sigma-Aldrich), MgCl2 to 1.5 mM (Promega), 1 unit of RedTaq DNA polymerase (Sigma-Aldrich) and Milli-Q water (Millipore) to a total volume of 20 μl. Reactions were carried out in a Master cycler S (Eppendorf) PCR machine. Products were run on a 1.5% 1×Tris-borate (TBE) agarose gels.


Once the sex PCR test was complete and analysed, the embryos were definitively labelled as either being genotypically male or female. The embryos were then opened via dissection and the gonads exposed for macroscopic analysis of gonadal development. The gonadal development of all control embryos was normal as expected. Control female embryo's showed typical asymmetric development that was characterised by a large left ovary and smaller regressing right gonad. Control male embryos all had typical bilateral testes. All female embryos from the siRNA knockdown groups (Groups 2-5) had normal gonadal development. In contrast, some male embryos from the siRNA knockdown groups showed varying degrees of female-like asymmetry at the macroscopic level of the gonads. The feminisation effect of the DMRT1-343-siRNA was characterised by an average or small-sized right testis and a larger feminised left gonad (Table 7). Feminisation was observed in a number of male embryos in Groups 2, 3 and 5 and resulted in an increase in the ratio of embryos with female-like gonads in these groups.









TABLE 7







DMRT1 embryo injection results.















%






Male:Female


siRNA dose
No.

% Male:Female
Macroscopic


and injection
embryos
No. viable at
Genotype
Gonad


route
injected
E10
(PCR sex test)
phenotype





Group 1
48
 48 (100%)
60:40
60:40


No injection






control






Group 2
51
39 (76%)
59:41
54:46


I.V.—100 pmol






Group 3
53
50 (94%)
54:46
46:54


I.V.—200 pmol






Group 4
81
33 (40%)
48:52
48:52


Amnion—100






pmol






Group 5
53
40 (75%)
33:67
28:72


Amnion—200






pmol









Gonads from both male and female embryos in each treatment groups were assessed for DMRT1 gene expression using quantitative RT-PCR analysis. Both the female and male gonads were pooled separately from each group and RNA was extracted for cDNA synthesis and qPCR analysis. The pooled gonads were added to 1 ml of Trizol and homogenised well by pipetting and vortexing at room temperature until all gonad tissue had dissolved. 200 μl of chloroform was added and mixed well by inverting the sample for 15 sec. The sample was then incubated at room temperature for 3 min and then centrifuged at 12000 g for 15 min at 4° C. The aqueous phase of the sample was then transferred to a new tube and then 500 l of isopropanol was added and mixed well by inversion. The mix was then incubated at room temperature for 10 min and then centrifuged at 12000 g for 10 min at 4° C. The supernatant was removed from the tube carefully, so as not to disturb the RNA pellet, and the pellet was then washed with 1 ml of 70% ethanol. The tube was then centrifuged at 7500 g for 5 min at 4° C. and the supernatant again was carefully removed and the RNA pellet was air dried at room temperature for 10 min. The RNA pellet was then resuspended in 25 μl of RNase-free water and the final concentration of RNA was determined using a NanoDrop ND-1000 Spectrophotometer (Thermo Scientific). RNA was reverse transcribed to complimentary DNA (cDNA) using the Promega Reverse Transcription kit (Promega). The reaction mix contained 1 μg of RNA, random hexamers (1 μl), dNTPs (2 μl), AMV reverse transcriptase (Promega) (0.5 μl) and nuclease free water added to a total reaction volume of 20 μl. The mix was incubated at 42° C. for 1 hour, followed by a 10 min incubation at 95° C. for enzyme inactivation.


cDNA was then used to quantify relative DMRT1 gene expression levels in the pooled male and female gonad samples from each treatment group. qPCR primers and probes were designed using Primer Express (Applied Biosystems) software and sequences are shown in Table 8. PCR's were set up in 20 μl reaction volumes that contained 2×TaqMan qRT PCR mastermix (Applied Biosystems), 1 μl of primer/probe mix, 1 μl of cDNA sample and made up to final volume with Nuclease free water (Promega). PCR cycling was performed at 95° C. for 1 min, followed by 40 cycles of 95° C. for 15 sec; 61° C. for 30 sec and; 68° C. for 30 sec. Ct values were obtained at a standard threshold value of 0.2 for all reactions. This threshold value corresponded to the midway point of the logarithmic phase of all amplification plots. Ct values were exported to Microsoft Excel for analysing relative gene expression using the comparative Ct method.









TABLE 8







Primer and probe sequences.








Sequence Name
Sequence 5′-3′





DMRT1 TaqMan MGB probe
CCATCCCTTTCATCTGCC



(SEQ ID NO: 3494)


DMRT1 Forward primer
TCAAGCCAGTCAGGAAAACAGT



(SEQ ID NO: 3495)


DMRT1 Reverse primer
TCATGGCATGGCGGTTCT



(SEQ ID NO: 3496)


18S rRNA TaqMan MGB probe
TGCTGGCACCAGACTTGCCCTC



(SEQ ID NO: 3497)


18S rRNA Forward primer
CGGCTACCACATCCAAGGAA



(SEQ ID NO: 3498)


18S rRNA Reverse primer
GCTGGAATTACCGCGGCT



(SEQ ID NO: 3499)









Relative levels of DMRT1 mRNA were compared with the chicken house keeping 18S rRNA species across all cDNA samples (FIG. 2). Quantitative RT-PCR analysis confirmed that DMRT1 mRNA expression was specifically reduced in all pooled groups of male embryos when compare to control Group 1. Almost 40% of DMRT1 gene expression knockdown was observed for Group 3 male embryos treated with the DMRT1-343-siRNA. It is interesting to note that Group 3 was also the group that resulted in the greatest degree of observed feminisation of male gonads at the macroscopic level.


It will be appreciated by persons skilled in the art that numerous variations and/or modifications may be made to the invention as shown in the specific embodiments without departing from the spirit or scope of the invention as broadly described. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.


All publications discussed and/or referenced herein are incorporated herein in their entirety.


The present application claims priority from U.S. 61/138,235 filed 17 Dec. 2008, the entire contents of which are incorporated by reference.


Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application.


REFERENCES



  • Clinton et al. (2001) British Poultry Science 42:134-138

  • Hori et al. (2000) Mol Biol Cell 11:3645-3660

  • Needleman and Wunsch (1970) J Mol Biol 48: 443-453

  • O'Neill et al. (2000) Dev Genes Evol 210:243-249

  • Raymond et al. (1999) Dev Biol 215:208-220

  • Smith et al. (1999) Nature 402:601-602

  • Smith et al. (2000) Nature 407: 319-320.

  • Taxman et al. (2006) BMC Biotechnol 6:7

  • Waterhouse et al. (1998) Proc Natl Acad Sci USA 95:13959-13964


Claims
  • 1-20. (canceled)
  • 21. A method of modifying the sex of an avian, the method comprising administering to an avian egg at least one isolated and/or exogenous RNA molecule comprising a double-stranded region (dsRNA) which reduces the level of a protein encoded by an R-spondin gene or Fox9 gene in the avian egg, wherein if the embryo of the egg is male the sex is altered to female following administration of the isolated and/or exogenous RNA molecule, and wherein the RNA molecule is administered to a non-cellular site of the egg and/or the egg is not electroporated.
  • 22. A method of modifying the sex of an avian, the method comprising administering to an avian egg at least one isolated and/or exogenous RNA molecule comprising a double-stranded region (dsRNA) comprising one or more of the sequence of nucleotides provided as SEQ ID NO's 11 to 3431 or a variant of any one or more thereof, wherein the isolated and/or exogenous RNA molecule does not comprise a sequence selected from: CCAGUUGUCAAGAAGAGCA (SEQ ID NO:254)GGAUGCUCAUUCAGGACAU (SEQ ID NO:369)CCCUGUAUCCUUACUAUAA (SEQ ID NO:474)GCCACUGAGUCUUCCUCAA (SEQ ID NO:530)CCAGCAACAUACAUGUCAA (SEQ ID NO:605)CCUGCGUCACACAGAUACU (SEQ ID NO:747)GGAGUAGUUGUACAGGUUG (SEQ ID NO:3432)GACUGGCUUGACAUGUAUG (SEQ ID NO:3433)AUGGCGGUUCUCCAUCCCU (SEQ ID NO:3434) or a variant of any one thereof, and wherein the RNA molecule is administered to a non-cellular site of the egg and/or the egg is not electroporated.
  • 23. A method of modifying the sex of an avian, the method comprising administering to an avian egg at least one isolated and/or exogenous RNA molecule comprising a double-stranded region (dsRNA) which reduces the level of a protein encoded by an β-catenin in the avian egg, wherein the RNA molecule is administered to a non-cellular site of the egg and/or the egg is not electroporated.
  • 24. The method of claim 21, wherein the non-cellular site is the air sac, yolk sac, amnionic cavity or chorion allantoic fluid.
  • 25. The method of claim 22, wherein the non-cellular site is the air sac, yolk sac, amnionic cavity or chorion allantoic fluid.
  • 26. The method of claim 23, wherein the non-cellular site is the air sac, yolk sac, amnionic cavity or chorion allantoic fluid.
  • 27. The method of claim 21, wherein the dsRNA is a siRNA or a shRNA.
  • 28. The method of claim 22, wherein the dsRNA is a siRNA or a shRNA.
  • 29. The method of claim 23, wherein the dsRNA is a siRNA or a shRNA.
  • 30. The method of claim 21, wherein the RNA molecule is administered by injection.
  • 31. The method of claim 22, wherein the RNA molecule is administered by injection.
  • 32. The method of claim 23, wherein the RNA molecule is administered by injection.
  • 33. The method of claim 21, wherein the avian is selected from the group consisting of chickens, ducks, turkeys, geese, bantams and quails.
  • 34. The method of claim 22, wherein the avian is selected from the group consisting of chickens, ducks, turkeys, geese, bantams and quails.
  • 35. The method of claim 23, wherein the avian is selected from the group consisting of chickens, ducks, turkeys, geese, bantams and quails.
  • 36. An avian produced using a method according of claim 21.
  • 37. An avian produced using a method according of claim 22.
  • 38. An avian produced using a method according of claim 23.
PCT Information
Filing Document Filing Date Country Kind 371c Date
PCT/AU2009/001627 12/16/2009 WO 00 12/21/2011
Provisional Applications (1)
Number Date Country
61138235 Dec 2008 US