Improve meat production and virus resistance by simultaneously editing multiple genes in livestock using Cas12iMax

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外文摘要：The clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated gene(Cas)system is continually optimized to achieve the most efficient gene editing effect.The Cas12iMax,a Cas12i variant,exhibits powerful DNA editing activity and enriches the gene editing toolbox.However,the application of Cas12iMax in large domestic animals has not yet been reported.To verify the efficiency and feasibility of multiple gene editing in large animals,we generated porcine fibroblasts with simultaneous knockouts of IGF2,ANPEP,CD163,and MSTN via Cas12iMax in one step.Phenotypically stable pigs were created through somatic cell nuclear transfer technology.They exhibited improved growth performance and muscle quality.Furthermore,we simultaneously edited three genes in bovine fibroblasts.A knockout of MSTN and PRNP was created and the amino acid Q-G in CD18 was precisely substituted.Meanwhile,no off-target phenomenon was observed by sum-type analysis or off-target detection.These results verified the effectiveness of Cas12iMax for gene editing in livestock animals and demonstrated the potential application of Cas12iMax in the field of animal trait improvement for agricultural production.

外文关键词：

Cas12iMaxgene editinglivestockmultiple trait improvement

作者：

Jilong Ren、Tang Hai、Yangcan Chen、Ke Sun、Zhiqiang Han、Jing Wang、Chongyang Li、Qingwei Wang、Leyun Wang、Huabing Zhu、Dawei Yu、Wei Li、Shanjiang Zhao

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作者单位：

State Key Laboratory of Animal Nutrition,Key Laboratory of Animal Genetics,Breeding and Reproduction of Ministry of Agriculture and Rural Affairs,Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193,China

Beijing Farm Animal Research Center,Institute of Zoology,Chinese Academy of Sciences,Beijing 100101,China

State Key Laboratory of Stem Cell and Reproductive Biology,Institute of Zoology,Chinese Academy of Sciences,Beijing 100101,China

Beijing Institute for Stem Cell and Regenerative Medicine,Beijing 100101,China

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基金：

国家重点研发计划国家重点研发计划国家重点研发计划国家重点研发计划国家自然科学基金National Key Research and Development Program ofNingxiaChina Agriculture Research System of MOF and MARAAgricultural Science and Technology Innovation Programproject from The Xinjiang Production and Construction Corps and Foundation of State Key Laboratory for Sheep Genetic Improve

项目编号：

2018YFE02011002021YFA08059052021YFA08057012022YFA1103101321025492021BEF02023CARS-36ASTIP-IAS062021ZD04

出版年：

2024

DOI：

10.1007/s11427-023-2407-0

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(3)

参考文献量33