title: Sleeping Beauty-mediated knockdown of sheep myostatin by RNA interference.
abstract: Myostatin is a negative regulator of skeletal muscle growth. Myostatin dysfunction therefore offers a strategy for promoting animal muscle growth in livestock production. Knockdown of myostatin was achieved by combining RNA interference and the Sleeping Beauty (SB) transposon system in sheep cells. Four targeting sites of sheep myostatin were designed and measured for myostatin silencing in sheep fetal fibroblasts by real-time PCR. The sh3 construct induced significant decrease of myostatin gene expression by 90% (P<0.05). Myostatin silencing induced by SB-mediated sh3 was further tested in stably transfected cells. SB transposition increased the integration frequency of genes into sheep genomes and mediated a more efficient myostatin knockdown than random integration of sh3. We suggest that SB-mediated shRNA provides a novel potential tool for gene knockdown in the donor cells of animal cloning.
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi 832003, China.
authors.forename: Shengwei
authors.initials: S
authors.lastname: Hu
authors.forename: Wei
authors.initials: W
authors.lastname: Ni
authors.forename: Wujiafu
authors.initials: W
authors.lastname: Sai
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Xudong
authors.initials: X
authors.lastname: Cao
authors.forename: Jun
authors.initials: J
authors.lastname: Qiao
authors.forename: Jinliang
authors.initials: J
authors.lastname: Sheng
authors.forename: Fei
authors.initials: F
authors.lastname: Guo
authors.forename: Chuangfu
authors.initials: C
authors.lastname: Chen
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005347:Fibroblasts / Q000502:physiology; D055785:Gene Knockdown Techniques / Q000379:methods*; D055435:Myostatin / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012756:Sheep; D014162:Transfection / Q000379:methods; D019895:Transposases / Q000235:genetics*
chemical_list: D055435:Myostatin; D034741:RNA, Small Interfering; D019895:Transposases