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Sci Rep:农科院杜立新等研究团队利用CRISPR/Cas9构建转基因绵羊

摘要 : 近日,国际学术权威刊物自然出版集团旗下子刊《Scientific Reports》杂志在线发表了中国农业科学院北京畜牧兽医研究所杜立新教授团队与中国农业大学王楚端研究组、暨南大学王晓刚研究组的联合团队合作的研究论文

 近日,国际学术权威刊物自然出版集团旗下子刊《Scientific Reports》杂志在线发表了中国农业科学院北京畜牧兽医研究所杜立新教授团队与中国农业大学王楚端研究组、暨南大学王晓刚研究组的联合团队合作的研究论文,研究利用CRISPR-Cas9系统成功生成了携带报告基因Rosa26的基因敲入转基因绵羊,该方法具有简单,设计灵活,效率高的特点。

绵羊(Sheep)是一种重要的经济动物,肌肉生长是绵羊育种的主要方向之一。Myostatin (MSTN)作为TGF-β超家族的一员,在肌肉生长中发挥负调控因子作用,其为改善家畜生长性能带来了巨大的希望。该团队在上述研究的基础上,利用CRISPR/Cas9系统高效构建出了myostatin (MSTN)基因突变羊,获得了MSTN基因突变羊5只,其中3只羊表现出臀部及后腿肌肉以及肩胸部肌肉肥大的双肌(DM)表型特征(附出生三月龄的转基因羊)。此项研究成果对于绵羊肌肉发育和产肉性能提高影响意义重大。

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原文链接:

Rosa26-targeted sheep gene knock-in via CRISPR-Cas9 system.

原文摘要:

Recent advances in our ability to design DNA binding factors with specificity for desired sequences have resulted in a revolution in genetic engineering, enabling directed changes to the genome to be made relatively easily. Technologies that facilitate specific and precise genome editing, such as knock-in, are critical for determining the functions of genes and for understanding fundamental biological processes. The CRISPR/Cas9 system has recently emerged as a powerful tool for functional genomic studies in mammals. Rosa26 gene can encode a non-essential nuclear RNA in almost all organizations, and become a hot point of exogenous gene insertion. Here, we describe efficient, precise CRISPR/Cas9-mediated Integration using a donor vector with tGFP sequence targeted in the sheep genomic Rosa26 locus. We succeeded in integrating with high efficiency an exogenous tGFP (turboGFP) gene into targeted genes in frame. Due to its simplicity, design flexibility, and high efficiency, we propose that CRISPR/Cas9-mediated knock-in will become a standard method for the generation transgenic sheep.

来源: Scientific Reports 浏览次数:0

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