文章摘要
Editing MSTN gene of Guangdong Xiaoerhuapig by using CRISPR/Cas9
  
DOI:10.16768/j.issn.1004-874X.2017.02.022
Author NameAffiliation
王 敏1,黄 翔1,石 翾1,刘小凤1,曾检华2,刘小红1,陈瑶生1,何祖勇1 1.中山大学生命科学学院/ 有害生物控制与资源利用国家重点实验室广东 广州 510006 2.广东壹号食品股份有限公司广东 广州 510620 
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Abstract:
      MSTN( Myostatin) gene functions as a negative regulator of muscle growth. Therefore,disrupting the expression of MSTN has been proved as an efficient strategy to improve the muscle mass in livestock and poultry. In this study,we edited the exons of MSTN gene to disrupt its expression in order to increase muscle production of Guangdong Xiaoerhua pig by using the cutting-edge genome editing technology—CRISPR/Cas9. We designed six guide RNAs targeting on loci across three exons of MSTN gene. Through T7E1 assay and TA cloning analysis,we found that three guide RNAs were capable of targeted cutting on MSTN gene efficiently, with the targeting efficiency ranging from 17.0% to 28.3%. The editing efficiency of gRNA1-1 and gRNA3-3 in PEF cells of Guangdong Xiaoerhua pig,were both up to 50%. Particularly,editing efficiency in the third exon was as high as 61.5%. This study provides basis to generate MSTN-edited Xiaoerhua pigs by somatic cell nuclease transfer(SCNT) in future.
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