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

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外文摘要：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 re-placement,and base editing,with high efficiency and specificity for biomedicine and agriculture.

外文关键词：

type Ⅰ CRISPR systemsDesulfovibrio vulgarisDvu type Ⅰ-C editordefine deletionlarge fragment replacementbase editing

作者：

Pan Li、Dingcai Dong、Fei Gao、Yuyang Xie、Honglin Huang、Siwei Sun、Zhao Ma、Cheng He、Jinsheng Lai、Xuguang Du、Sen Wu

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作者单位：

State Key Laboratory of Animal Biotech Breeding,College of Biological Sciences,National Engineering Laboratory for Animal Breeding,Frontiers Science Center for Molecular Design Breeding,China Agricultural University,Beijing 100193,China

College of Veterinary Medicine,China Agricultural University,Beijing 100193,China

Sanya Institute of China Agricultural University,Sanya 572025,China

State Key Laboratory of Maize Bio-breeding,National Maize Improvement Center,Department of Plant Genetics and Breeding,China Agricultural University,Beijing 100193,China

Frontiers Science Center for Molecular Design Breeding,China Agricultural University,Beijing 100193,China

Center for Crop Functional Genomics and Molecular Breeding,China Agricultural University,Beijing 100193,China

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出版年：

2024

DOI：

10.1007/s11427-023-2682-5

中国科学:生命科学(英文版)

中国科学院

中国科学:生命科学(英文版)

CSTPCD

影响因子：0.806

ISSN：1674-7305

年,卷(期)：2024.67(11)