Versatile and efficient mammalian genome editing with Type I-C CRISPR System of Desulfovibrio vulgaris.

IF 8 2区 生物学 Q1 BIOLOGY Science China Life Sciences Pub Date : 2024-11-01 Epub Date: 2024-08-07 DOI:10.1007/s11427-023-2682-5
Pan Li, Dingcai Dong, Fei Gao, Yuyang Xie, Honglin Huang, Siwei Sun, Zhao Ma, Cheng He, Jinsheng Lai, Xuguang Du, Sen Wu
{"title":"Versatile and efficient mammalian genome editing with Type I-C CRISPR System of Desulfovibrio vulgaris.","authors":"Pan Li, Dingcai Dong, Fei Gao, Yuyang Xie, Honglin Huang, Siwei Sun, Zhao Ma, Cheng He, Jinsheng Lai, Xuguang Du, Sen Wu","doi":"10.1007/s11427-023-2682-5","DOIUrl":null,"url":null,"abstract":"<p><p>CRISPR-Cas tools for mammalian genome editing typically rely on single Cas9 or Cas12a proteins. While type I CRISPR systems in Class I may offer greater specificity and versatility, they are not well-developed for genome editing. Here, we present an alternative type I-C CRISPR system from Desulfovibrio vulgaris (Dvu) for efficient and precise genome editing in mammalian cells and animals. We optimized the Dvu type I-C editing complex to generate precise deletions at multiple loci in various cell lines and pig primary fibroblast cells using a paired PAM-in crRNA strategy. These edited pig cells can serve as donors for generating transgenic cloned piglets. The Dvu type I-C editor also enabled precise large fragment replacements with homology-directed repair. Additionally, we adapted the Dvu-Cascade effector for cytosine and adenine base editing, developing Dvu-CBE and Dvu-ABE systems. These systems efficiently induced C-to-T and A-to-G substitutions in human genes without double-strand breaks. Off-target analysis confirmed the high specificity of the Dvu type I-C editor. Our findings demonstrate the Dvu type I-C editor's potential for diverse mammalian genome editing applications, including deletions, fragment replacement, and base editing, with high efficiency and specificity for biomedicine and agriculture.</p>","PeriodicalId":21576,"journal":{"name":"Science China Life Sciences","volume":" ","pages":"2471-2487"},"PeriodicalIF":8.0000,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Science China Life Sciences","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1007/s11427-023-2682-5","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/8/7 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"BIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

CRISPR-Cas tools for mammalian genome editing typically rely on single Cas9 or Cas12a proteins. While type I CRISPR systems in Class I may offer greater specificity and versatility, they are not well-developed for genome editing. Here, we present an alternative type I-C CRISPR system from Desulfovibrio vulgaris (Dvu) for efficient and precise genome editing in mammalian cells and animals. We optimized the Dvu type I-C editing complex to generate precise deletions at multiple loci in various cell lines and pig primary fibroblast cells using a paired PAM-in crRNA strategy. These edited pig cells can serve as donors for generating transgenic cloned piglets. The Dvu type I-C editor also enabled precise large fragment replacements with homology-directed repair. Additionally, we adapted the Dvu-Cascade effector for cytosine and adenine base editing, developing Dvu-CBE and Dvu-ABE systems. These systems efficiently induced C-to-T and A-to-G substitutions in human genes without double-strand breaks. Off-target analysis confirmed the high specificity of the Dvu type I-C editor. Our findings demonstrate the Dvu type I-C editor's potential for diverse mammalian genome editing applications, including deletions, fragment replacement, and base editing, with high efficiency and specificity for biomedicine and agriculture.

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
利用普通脱硫弧菌的 I-C 型 CRISPR 系统进行多功能、高效的哺乳动物基因组编辑。
用于哺乳动物基因组编辑的 CRISPR-Cas 工具通常依赖单个 Cas9 或 Cas12a 蛋白。虽然I类CRISPR系统可能具有更高的特异性和多功能性,但它们在基因组编辑方面并不发达。在这里,我们介绍了一种来自Desulfovibrio vulgaris(Dvu)的I-C型CRISPR系统,用于在哺乳动物细胞和动物中进行高效、精确的基因组编辑。我们对 Dvu I-C 型编辑复合物进行了优化,利用成对的 PAM-in crRNA 策略在各种细胞系和猪原代成纤维细胞中生成多个位点的精确缺失。这些经过编辑的猪细胞可作为供体,用于产生转基因克隆仔猪。Dvu I-C 型编辑器还能通过同源定向修复实现大片段的精确替换。此外,我们还将 Dvu 级联效应器用于胞嘧啶和腺嘌呤碱基编辑,开发出了 Dvu-CBE 和 Dvu-ABE 系统。这些系统能有效地诱导人类基因中的 C 到 T 和 A 到 G 的置换,而不会发生双链断裂。脱靶分析证实了 Dvu I-C 型编辑器的高度特异性。我们的研究结果表明,Dvu I-C 型编辑器可用于多种哺乳动物基因组编辑应用,包括缺失、片段置换和碱基编辑,在生物医学和农业领域具有高效率和特异性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
CiteScore
15.10
自引率
8.80%
发文量
2907
审稿时长
3.2 months
期刊介绍: Science China Life Sciences is a scholarly journal co-sponsored by the Chinese Academy of Sciences and the National Natural Science Foundation of China, and it is published by Science China Press. The journal is dedicated to publishing high-quality, original research findings in both basic and applied life science research.
期刊最新文献
Exaptation of pectoral fins for olfaction in the spiny red gurnard (Chelidonichthys spinosus) through an ancient receptor. Genomic analysis of modern maize inbred lines reveals diversity and selective breeding effects. Genome editing technology and medical applications. Dual activation of soybean resistance against Phytophthora sojae by pectin lyase and degraded pectin oligosaccharides. Selenium metabolism and selenoproteins function in brain and encephalopathy.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1