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Updated: Sep 15, 2025

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获得的CRISPR-Cas12f同类驱动RNA引导的转录

Florian T Hoffmann¹, Tanner Wiegand^1,2, Adriana I Palmieri¹

¹Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, USA.

bioRxiv : the preprint server for biology

|July 15, 2025

概括

研究人员使用dCas12f-sigmaE复合体在细菌中发现了一种新的RNA引导的基因激活机制. 这个系统绕过了传统的促进者,为细菌转录调节和潜在的合成生物学应用提供了新的见解.

科学领域:

分子生物学分子生物学
细菌遗传学细菌遗传学
基因规则基因规则

背景情况:

细菌转录启动通常需要RNA聚合酶促进体识别的西格玛因子.
细菌体拥有众多未表征的细胞外功能西格玛因子 (σ^E),这表明它们具有复杂的调节作用.
现有的知识转录启动在E. coli专注于定义良好的西格玛因子函数.

研究的目的:

调查细菌体中专门的s^E因子未知的作用.
发现细菌基因激活的新机制.
探索dCas12f-σE复合体在可编程基因表达中的潜力.

主要方法:

在E中对dCas12f和s^E同类的选. 大肠杆菌.
使用RNA免疫沉降测序 (RIP-seq) 和染色体免疫沉降测序 (ChIP-seq).
共同表达实验以评估RNA聚合酶全酶招募.

主要成果:

确定了dCas12f-σ^E复合体,能够引导RNA丰富和DNA标结合.
通过dCas12f-gRNA-σ^E三元复合体证明了RNA聚合酶全酶的可编程招募.
实现了独立于促进子动机的强烈基因表达,转录起始点由dCas12f介导的R循环决定.

结论:

发现了一种前所未有的RNA引导转录 (RGT) 机制.
这种机制利用dCas12f和s^E来激活无促进子的基因.
这些发现为基因调节提供了一个新的范式,反映了CRISPRa技术的各个方面.

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