Impact Factor:13.214

DOI number:10.1186/s13059-016-1144-4

Journal:Genome Biology (B刊)

Abstract: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.

Co-author:Liu X,Chen LL,Li Q,Cui CC,Liu Xu,Zhang JC

First Author:Gao YP#,Wu HB#,Wang YS#

Indexed by:Journal paper

Correspondence Author:Zhang Y*

Volume:18

Page Number:13

Translation or Not:no

Date of Publication:2017-02-01

Included Journals:SCI

Links to published journals:https://genomebiology.biomedcentral.com/articles/10.1186/s13059-016-1144-4