title: Myostatin knockout in bovine fetal fibroblasts by using TALEN.
abstract: Myostatin (MSTN) can negatively regulate the growth and development of skeletal muscle, and mutations of bovine MSTN gene can cause a "double-muscle" feature. To knock out MSTN gene in bovine fetal fibroblast by transcription activator-like effector nucleases (TALENs) and obtain MSTN knockout cell lines, we constructed one pair of MSTN-TALEN vector and transfected into bovine fetal fibroblast cells by PEI and electroporation. Sequencing results demonstrated that TALEN was available for MSTN knockout. T7 endonuclease 1 (T7E1) was used for the detection of mutation efficiency. The results indicated that knockout efficiency of electroporation transfection was 20.4%, and 10 MSTN(+/-) and MSTN(-/-) cell colonies were obtained via limiting dilution method. The deletion number of nucleotides ranged from 1 to 20, and some of them were frameshift mutation, which could provide the possibility in production of MSTN knockout cattle in the future.
authors: College of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors: Cuicui
authors: C
authors: Yang
authors: College of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors: Huili
authors: H
authors: Tong
authors: College of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors: Xinghong
authors: X
authors: Ma
authors: College of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors: Wei
authors: W
authors: Du
authors: College of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors: Dan
authors: D
authors: Liu
authors: College of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors: Yu
authors: Y
authors: Yang
authors: College of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors: Yunqin
authors: Y
authors: Yan
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D003851:Deoxyribonucleases / Q000378:metabolism; D018274:Electroporation; D005347:Fibroblasts / Q000378:metabolism*; D055786:Gene Knockout Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D009154:Mutation; D055435:Myostatin / Q000172:deficiency / Q000235:genetics*
chemical_list: D055435:Myostatin; D003851:Deoxyribonucleases