title: Cloning and functional analysis of sheep U6 promoters.
abstract: Gene silencing mediated by small interfering RNA has become a powerful biological tool for the regulation of gene expression. In order to develop an effective short hairpin RNA (shRNA) expression vector, specifically for use in sheep species, we have identified two sheep U6 promoters based on the highly conserved polymerase III promoter elements. Promoter activity was measured by U6 promoter-driven shRNA to suppress enhanced green fluorescent protein (EGFP) expression. The knock down assay demonstrated that the two sheep U6 promoters and mouse U6 promoter induced a similar level of EGFP knockdown. These results suggest that the two sheep U6 promoters could efficiently drive shRNA expression for gene silencing and may have applications in RNAi-based sheep research.
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi, China.
authors.forename: Shengwei
authors.initials: S
authors.lastname: Hu
authors.forename: Wei
authors.initials: W
authors.lastname: Ni
authors.forename: Wureli
authors.initials: W
authors.lastname: Hazi
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Na
authors.initials: N
authors.lastname: Zhang
authors.forename: Ren
authors.initials: R
authors.lastname: Meng
authors.forename: Chuangfu
authors.initials: C
authors.lastname: Chen
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods; D005347:Fibroblasts; D055785:Gene Knockdown Techniques / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D012342:RNA, Small Nuclear / Q000235:genetics*; D012756:Sheep / Q000235:genetics*; D014162:Transfection
chemical_list: D012342:RNA, Small Nuclear; C405486:U6 small nuclear RNA; D049452:Green Fluorescent Proteins