Versatile and robust genome editing with Streptococcus thermophilus CRISPR1-Cas9

Author:

Agudelo DanielORCID,Carter SophieORCID,Velimirovic MinjaORCID,Duringer AlexisORCID,Rivest Jean-FrançoisORCID,Levesque SébastienORCID,Loehr JeremyORCID,Mouchiroud MathildeORCID,Cyr DenisORCID,Waters Paula J.ORCID,Laplante MathieuORCID,Moineau SylvainORCID,Goulet AdelineORCID,Doyon YannickORCID

Abstract

Targeting definite genomic locations using CRISPR-Cas systems requires a set of enzymes with unique protospacer adjacent motif (PAM) compatibilities. To expand this repertoire, we engineered nucleases, cytosine base editors, and adenine base editors from the archetypal Streptococcus thermophilus CRISPR1-Cas9 (St1Cas9) system. We found that St1Cas9 strain variants enable targeting to five distinct A-rich PAMs and provide a structural basis for their specificities. The small size of this ortholog enables expression of the holoenzyme from a single adeno-associated viral vector for in vivo editing applications. Delivery of St1Cas9 to the neonatal liver efficiently rewired metabolic pathways, leading to phenotypic rescue in a mouse model of hereditary tyrosinemia. These robust enzymes expand and complement current editing platforms available for tailoring mammalian genomes.

Funder

Canadian Institutes of Health Research

Banting Research Foundation

French National Research Agency

Resource for Biocomputing, Visualization, and Informatics

National Institutes of Health

NIH

Fonds de la recherche du Québec-Santé

Vanier Canada graduate scholarship

Frederick Banting and Charles Best Canada graduate scholarship

Fondation du Grand défi Pierre Lavoie

Publisher

Cold Spring Harbor Laboratory

Subject

Genetics(clinical),Genetics

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