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

IF 8 2区 生物学 Q1 BIOLOGY Science China Life Sciences Pub Date : 2024-11-01 Epub Date: 2024-08-02 DOI:10.1007/s11427-023-2572-6
Lin Zhang, Shi-Meng Cao, Hao Wu, Meng Yan, Jinsong Li, Ling-Ling Chen
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Abstract

The growing variety of RNA classes, such as mRNAs, lncRNAs, 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 lncRNA, 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.

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CRISPR/RfxCas13d 介导的小鼠胚胎发育中高效 RNA 敲除策略。
越来越多的 RNA 种类,如 mRNA、lncRNA 和 circRNA,在发育过程和各种病理生理状况中发挥着关键作用。然而,由于缺乏直接影响 RNA 水平的持久有效的策略,我们对 RNA 在活生物体中功能的理解仍然有限。在这项研究中,我们将 CRISPR-RfxCas13d 系统与类精干细胞介导的半克隆技术结合起来,从而抑制了不同 RNA 种类的表达。我们采用这种方法干扰了三种 RNA 分子的表达:Sfmbt2 mRNA、Fendrr lncRNA 和 circMan1a2(2,3,4,5,6)。结果证实了这些 RNA 在胚胎发育中的关键作用,因为它们的缺失会导致可观察到的表型,包括胚胎致死、胚胎发育延迟和胚胎吸收。总之,我们的方法为沉默生物体中的特定 RNA 靶标提供了一个有效的工具包,而无需引入基因改变。
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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.
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