首页|A CRISPR/RfxCas13d-mediated strategy for efficient RNA knockdown in mouse embryonic development

A CRISPR/RfxCas13d-mediated strategy for efficient RNA knockdown in mouse embryonic development

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The growing variety of RNA classes,such as mRNAs,IncRNAs,and circRNAs,plays pivotal roles in both developmental processes and various pathophysiological conditions.Nonetheless,our comprehension of RNA functions in live organisms remains limited due to the absence of durable and effective strategies for directly influencing RNA levels.In this study,we combined the CRISPR-RfxCas13d system with sperm-like stem cell-mediated semi-cloning techniques,which enabled the suppressed expression of different RNA species.This approach was employed to interfere with the expression of three types of RNA molecules:Sfmbt2 mRNA,Fendrr IncRNA,and circMan1a2(2,3,4,5,6).The results confirmed the critical roles of these RNAs in embryonic development,as their loss led to observable phenotypes,including embryonic lethality,delayed embryonic development,and embryo resorption.In summary,our methodology offers a potent toolkit for silencing specific RNA targets in living organisms without introducing genetic alterations.

sperm-like stem cellsemi-cloned technologyCRISPR-RfxCas1 3d I RNA knockdownembryonic development

Lin Zhang、Shi-Meng Cao、Hao Wu、Meng Yan、Jinsong Li、Ling-Ling Chen

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State Key Laboratory of Cell Biology,Shanghai Key Laboratory of Molecular Andrology,Shanghai Institute of Biochemistry and Cell Biology,Center for Excellence in Molecular Cell Science,Chinese Academy of Sciences,University of Chinese Academy of Sciences,Shanghai 200031,China

Key Laboratory of RNA Innovation,Science and Engineering,CAS Center for Excellence in Molecular Cell Science,Shanghai Institute of Biochemistry and Cell Biology,University of Chinese Academy of Sciences,Chinese Academy of Sciences,Shanghai 200031,China

Key Laboratory of Systems Health Science of Zhejiang Province,School of Life Science,Hangzhou Institute for Advanced Study,University of Chinese Academy of Sciences,Hangzhou 310024,China

School of Life Science and Technology,ShanghaiTec

School of Life Science and Technology,ShanghaiTech University,Shanghai 2

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2024

10.1007/s11427-023-2572-6

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)