title: Identification and analysis of genetic variations in pri-miRNAs expressed specifically or at a high level in sheep skeletal muscle.
abstract: MicroRNAs (miRNAs) are key regulators in miRNA-mediated gene regulatory networks and play important roles in many biological processes, such as growth and development of mammals. In this study, we used microarrays to detect 261 miRNAs that are expressed in sheep skeletal muscle. We found 22 miRNAs that showed high levels of expression and equated to 89% of the total miRNA. Genetic variations in these 22 pri-miRNAs were further investigated using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and sequencing. A total of 49 genetic variations, which included 41 single nucleotide polymorphisms (SNPs) and 8 deletions/insertions, were identified in four sheep breeds. Three variations were further researched in a larger sample set, including five sheep breeds with different meat production performances. We found that the genotype and allele frequencies of the CCC deletion/insertion in pri-miR-133a were significantly related to the sheep meat production trait. Finally, cell assays and quantitative reverse transcription PCR (qRT-PCR) were employed to investigate the effect of pri-miRNA genetic variation on the miRNA biogenesis process. The results confirmed that genetic variations can influence miRNA biogenesis and increase or decrease the levels of mature miRNAs, in accordance with the energy and stability change of hair-pin secondary structures. Our findings will help to further the understanding of the functions of genetic variations in sheep pri-miRNAs in skeletal muscle growth and development.
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, 832003, China; Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China.
authors.forename: Limin
authors.initials: L
authors.lastname: Wang
authors.affiliation: Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhou
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, 832003, China; Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China.
authors.forename: Guangchao
authors.initials: G
authors.lastname: Song
authors.affiliation: Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China.
authors.forename: Min
authors.initials: M
authors.lastname: Shen
authors.affiliation: Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China.
authors.forename: Shangquan
authors.initials: S
authors.lastname: Gan
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, 832003, China; Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China.
authors.forename: Guoqing
authors.initials: G
authors.lastname: Shi
mesh_terms: D000483:Alleles; D000818:Animals; D005787:Gene Frequency; D014644:Genetic Variation; D005838:Genotype; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D020641:Polymorphism, Single Nucleotide; D012322:RNA Precursors / Q000378:metabolism; D020871:RNA Stability; D060888:Real-Time Polymerase Chain Reaction; D017423:Sequence Analysis, RNA; D012756:Sheep
chemical_list: D035683:MicroRNAs; D012322:RNA Precursors