Detection of Lethality Gene for Improved Fertility in Mammals

Abstract

Oligonucleic acid molecules comprising a SNP site at a position corresponding to position 7480 of the bovine signal transducer and activator of transcription (STAT5A) coding sequence (SEQ ID NO: 1). Also disclosed are an array or a kit comprising the same, a method for detecting the SNPs, a method for progeny testing of mammals, a method for increasing human and non-human mammal pregnancy rate in natural and artificial reproduction processes. Further provided are cattle breeding methods for improved milk production traits.

1 . An isolated single or double stranded oligonucleotide molecule comprising a polymorphic site at a position corresponding to position 7480 of exon 8 of bovine Signal Transducer and Activator 5A (STAT5A) coding sequence (SEQ ID NO: 1), wherein position 7480 is cytosine or guanine, and at least about 9 contiguous nucleotides of SEQ ID NO: 1 adjacent to the polymorphic site.  
     
     
         2 . The oligonucleotide molecule according to  claim 1 , which comprises at least about 15 contiguous nucleotides adjacent to the polymorphic site.  
     
     
         3 . The oligonucleotide molecule according to  claim 2 , which comprises at least about 20 contiguous nucleotides adjacent to the polymorphic site.  
     
     
         4 . The ioligonucleotide molecule according to  claim 1 , which comprises not more than about 150 nucleotides.  
     
     
         5 . The oligonucleotide molecule according to  claim 1 , which comprises not more than about 100 nucleotides.  
     
     
         6 . The oligonucleotide molecule according to  claim 1 , which comprises not more than about 50 nucleotides.  
     
     
         7 . The oligonucleotide molecule according to  claim 1 , wherein the polymorphic site is within 4 nucleotides of the center of the oligonucleotide molecule.  
     
     
         8 . The oligonucleotide molecule according to  claim 7 , wherein the polymorphic site is at the center of the oligonucleotide molecule.  
     
     
         9 . The oligonucleotide molecule according to  claim 1 , wherein the polymorphic site is at the 3′-end of the oligonucleotide molecule.  
     
     
         10 . An array of nucleic acid molecules comprising the isolated oligonucleotide molecules according to  claim 1  supported on a substrate.  
     
     
         11 . The array according to  claim 10 , further comprising one or more markers in linkage disequilibrium with the polymorphic site.  
     
     
         12 . A kit comprising a nucleic acid molecule of  claim 1 , and a suitable container.  
     
     
         13 . A method for detecting single nucleotide polymorphism (SNP) on STAT5A coding region in an animal cell, the method comprising determining the identity of a nucleotide at a position corresponding to position 7480 of exon 8 of bovine STAT5A coding sequence (SEQ ID NO: 1) of the cell.  
     
     
         14 . The method according to  claim 13 , wherein the animal is a mammalian, avian or aquatic species.  
     
     
         15 . The method according to  claim 13 , wherein the animal cell is an adult cell, an embryo cell, a sperm, an egg, a fertilized egg, or a zygote.  
     
     
         16 . A method according to  claim 14 , wherein the animal is a mammal.  
     
     
         17 . A method according to  claim 13 , wherein the mammal is bovine.  
     
     
         18 . A method according to  claim 13 , wherein the identity of the nucleotide is determined by sequencing a nucleic acid molecule comprising a position corresponding to position 7480 of exon 8 of bovine STAT5A coding sequence, or a relevant fragment thereof, isolated from the cell.  
     
     
         19 . A method according to  claim 18 , wherein the nucleic acid molecule is isolated from the cell via amplification by the polymerase chain reaction (PCR) of genomic DNA of the cell, or by RT-PCR of the mRNA of the cell.  
     
     
         20 . A method according to  claim 13 , wherein the identity of the nucleotide is determined by hybridizing a suitable probe to a preparation comprising the nucleic acid from the cell.  
     
     
         21 . A method according to  claim 20 , wherein the probe is labeled with a detectable label.  
     
     
         22 . A method according to  claim 13 , wherein the identity of the nucleotide is determined by an invasive signal amplification assay.  
     
     
         23 . A method according to  claim 13 , wherein the identity of the nucleotide of both copies of the coding sequence of the cell is determined.  
     
     
         24 . A method according to  claim 12 , wherein the identity of the nucleotide of both copies of the sequence is determined based on genotypes of the parent of the cell, genotypes of the daughter of the cell, or both.  
     
     
         25 . A method for determining whether an individual animal is suitable as a gamete donor for natural mating, artificial insemination or in vitro fertilization, the method comprising determining allele identify of the SNP site according to  claim 13 , or of an allele in linkage disequilibrium with the SNP site, and selecting as gamete donor an individual whose genotype is homozygous with regard to C allele at the SNP site, or homozygous with regard to an allele in linkage disequilibrium with the C allele.  
     
     
         26 . The method according to  claim 25 , wherein the animal is selected from the group consisting of cattle, swine, equine, dog, sheep and goat.  
     
     
         27 . The method according to  claim 25 , wherein the animal is human.  
     
     
         28 . A method of selecting an embryo for planting in a uterus, the method comprising determining identify of the nucleotide at a position corresponding to position 7480 of bovine STAT5A (SEQ ID NO: 1) of the embryo while preserving the viability of the embryo, and selecting for planting only an embryo whose genotype is CC homozygous at the position.  
     
     
         29 . The method according to  claim 28 , wherein the animal is an animal selected from the group consisting of cattle, swine, equine, dog, sheep and goat.  
     
     
         30 . The method according to  claim 29 , wherein multiple ovulation and embryo transfer (MOET) is used to generate multiple fertilized eggs.  
     
     
         31 . The method according to  claim 28 , wherein the animal is human.  
     
     
         32 . A method for increasing successful pregnancy rate of a non-human animal, comprising selecting a male or a female mammal for breeding purposes that are CC homozygous at a position correspond to position 7480 of exon 8 of bovine STAT5A gene (SEQ ID NO: 1).  
     
     
         33 . A method according to  claim 32 , wherein both male and female parents are CC homozygous.  
     
     
         34 . The method according to  claim 33 , wherein the female mammal is in vitro fertilized.  
     
     
         35 . A method for increasing pregnancy rate and reducing multiple pregnancy rate in a human ART procedure, the method comprising genotyping, via pre-implantation genetic diagnosis, the genotype of embryos to be planted with regard to the nucleotide corresponding to position 7480 of exon 8 of bovine STAT5A gene (SEQ ID NO: 1), and planting not more than 3 embryos which are homozygous CC with regard to the position.  
     
     
         36 . A method for determining whether an individual dairy cattle is suitable as a gamete donor for natural mating, artificial insemination or in vitro fertilization, the method comprising determining allele identify of the SNP site according to  claim 13 , or of an allele in linkage disequilibrium with the SNP site, and selecting as gamete donor an individual whose genotype is heterozygous at the SNP site, or heterozygous with regard to a locus in linkage disequilibrium with the SNP site.  
     
     
         37 . The method according to  claim 36 , wherein an individual having CC genotype at the SNP site is selected to mate with an individual with a CG genotype.  
     
     
         38 . The method according to  claim 37 , wherein gametes from the CG individual and the CC individual are used in artificial insemination, or in in vitro fertilization, or in multiple ovulation and embryo transfer procedure.  
     
     
         39 . A method of selecting a dairy cattle embryo for planting in a uterus, the method comprising determining identify of the nucleotide at a position corresponding to position 7480 of bovine STAT5A (SEQ ID NO: 1) of the embryo while preserving the viability of the embryo, and selecting for planting only an embryo whose genotype is CG heterozygous at the position.