Generation of B Cell-Deficient Pigs by Highly Efficient CRISPR/Cas9-Mediated Gene Targeting. Issue 8 (20th August 2015)

• Record Type: Journal Article
• Authors: Chen, Fengjiao; Wang, Ying; Yuan, Yilin; Zhang, Wei; Ren, Zijian; Jin, Yong; Liu, Xiaorui; Xiong, Qiang; Chen, Qin; Zhang, Manling; Li, Xiaokang; Zhao, Lihua; Li, Ze; Wu, Zhaoqiang; Zhang, Yanfei; Hu, Feifei; Huang, Juan; Li, Rongfeng; Dai, Yifan
• Is Part Of: Journal of genetics and genomics. Volume 42:Issue 8(2015:Aug.)
• Page Start: 437
• Page End: 444
• Publication Date: 2015-08-20
• Subjects: Genetics -- Periodicals; Genomics -- Periodicals; 576.505
• DOI: 10.1016/j.jgg.2015.05.002
• Languages: English
• ISSNs: 1673-8527

Abstract

<abstract xml:lang="en" abstract-type="author" id="abs0010"> <title id="sectitle0010">Abstract</title> <sec> <p id="abspara0010">Generating B cell-deficient mutant is the first step to produce human antibody repertoires in large animal models. In this study, we applied the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system to target the J_H region of the pig <italic>IgM</italic> heavy chain gene which is crucial for B cell development and differentiation. Transfection of IgM-targeting Cas9 plasmid in primary porcine fetal fibroblasts (PFFs) enabled inducing gene knock out (KO) in up to 53.3% of colonies analyzed, a quarter of which harbored biallelic modification, which was much higher than that of the traditional homologous recombination (HR). With the aid of somatic cell nuclear transfer (SCNT) technology, three piglets with the biallelic <italic>IgM</italic> heavy chain gene mutation were produced. The piglets showed no antibody-producing B cells which indicated that the biallelic mutation of the <italic>IgM</italic> heavy chain gene effectively knocked out the function of the <italic>IgM</italic> and resulted in a B cell-deficient phenotype. Our study suggests that the CRISPR/Cas9 system combined with SCNT technology is an efficient genome-editing approach in pigs.</p> </sec> </abstract>