利用TALEN技术在牛胎儿成纤维细胞中敲除Myostatin基因

doi:10.3724/SP.J.1005.2014.0685

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HEREDITAS(Beijing) ›› 2014, Vol. 36 ›› Issue (7): 685-690.doi: 10.3724/SP.J.1005.2014.0685

• Research Articles • Previous Articles Next Articles

Myostatin knockout in bovine fetal fibroblasts by using TALEN

Cuicui Yang, Huili Tong, Xinghong Ma, Wei Du, Dan Liu, Yu Yang, Yunqin Yan

College of Life Science, Northeast Agricultural University, Harbin 150030, China

Received:2013-12-16 Online:2014-07-20 Published:2014-06-23

Abstract

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.

Key words: TALENs, bovine, knockout, transfection

Cuicui Yang, Huili Tong, Xinghong Ma, Wei Du, Dan Liu, Yu Yang, Yunqin Yan. Myostatin knockout in bovine fetal fibroblasts by using TALEN[J]. HEREDITAS(Beijing), 2014, 36(7): 685-690.

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Myostatin knockout in bovine fetal fibroblasts by using TALEN

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