影响因子：13.214

DOI码：10.1186/s13059-016-1144-4

发表刊物：Genome Biology (B刊)

摘要：Background The CRISPR-Cas9 system is a widely utilized platform for transgenic animal production in various species, although its off-target effects should be addressed. Several applications of this tool have been proposed in model animals but remain insufficient for transgenic livestock production. Results Here, we report the first application of single Cas9 nickase (Cas9n) to induce gene insertion at a selected locus in cattle. We identify the main binding sites of a catalytically inactive Cas9 (dCas9) protein in bovine fetal fibroblast cells (BFFs) with chromatin immunoprecipitation sequencing (ChIP-seq). Subsequently, we demonstrate that a single Cas9n-induced single-strand break can stimulate the insertion of the natural resistance-associated macrophage protein-1 (NRAMP1) gene with reduced, but still considerable, off-target effects. Through somatic cell nuclear transfer, we finally obtain transgenic cattle with increased resistance to tuberculosis. Conclusions Our results contribute to the development of CRISPR-Cas9 system for agriculture applications.

合写作者：Liu X,Chen LL,Li Q,Cui CC,Liu Xu,Zhang JC

第一作者：Gao YP#,Wu HB#,Wang YS#

论文类型：期刊论文

通讯作者：Zhang Y*

卷号：18

页面范围：13

是否译文：否

发表时间：2017-02-01

收录刊物：SCI

发布期刊链接：https://genomebiology.biomedcentral.com/articles/10.1186/s13059-016-1144-4