Highly efficient CRISPR‐mediated base editing for the gut Bacteroides spp. with pnCasBS‐CBE

Author:

Liang Jing1,Tan Yang1

Affiliation:

1. Institute of Synthetic Biology Shenzhen Institute of Advanced Technology Chinese Academy of Sciences Shenzhen China

Abstract

AbstractBacteroidales are the most abundant order of bacteria in the healthy human gut and have the potential as a therapeutic agent. We constructed a pnCasBS‐CBE system for base editing in the Bacteroides thetaiotaomicron to expand their genetic toolkit, which is able to efficiently convert a C:G to a T:A in the genome. As a functional proof‐of‐concept, we used the pnCasBS‐CBE system to successfully introduce nonsynonymous mutation and stop codons to the genes involved in carbohydrate metabolism. The system also allowed for multiplexed gene editing with a single plasmid, enabling efficient editing of up to four genes in a single experiment. Furthermore, the pnCasBS‐CBE editing system was validated and successfully applied in four other non‐model gut Bacteroides species for genome editing. An unbiased genome‐wide SNPs analysis indicated that the pnCasBS‐CBE system showed high fidelity and applicability. Thus, this study provides a powerful clustered regularly interspaced short palindromic repeats (CRISPR)‐mediated genome editing toolbox for functional genomic analysis in Bacteroidales.

Funder

Shenzhen Institutes of Advanced Technology Innovation Program for Excellent Young Researchers

China Postdoctoral Science Foundation

Publisher

Wiley

Subject

Molecular Medicine,Applied Microbiology and Biotechnology,General Medicine

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3