Suppr超能文献

通过条件稳定的 CRISPR-Cas13b 编辑器进行位点特异性 mA 擦除。

Site-Specific m A Erasing via Conditionally Stabilized CRISPR-Cas13b Editor.

机构信息

Department of Chemistry, Case Western Reserve University, 2080 Adelbert Rd, Cleveland, OH 44106, USA.

出版信息

Angew Chem Int Ed Engl. 2023 Oct 23;62(43):e202309291. doi: 10.1002/anie.202309291. Epub 2023 Sep 15.

Abstract

N6-methyladenosine (m A) on RNAs plays an important role in regulating various biological processes and CRIPSR technology has been employed for programmable m A editing. However, the bulky size of CRISPR protein and constitutively expressed CRISPR/RNA editing enzymes can interfere with the native function of target RNAs and cells. Herein, we reported a conditional m A editing platform (FKBP*-dCas13b-ALK) based on a ligand stabilized dCas13 editor. The inducible expression of this m A editing system was achieved by adding or removing the Shield-1 molecule. We further demonstrated that the targeted recruitment of dCas13b-m A eraser fusion protein and site-specific m A erasing were achieved under the control of Shield-1. Moreover, the release and degradation of dCas13b fusion protein occurred faster than the restoration of m A on the target RNAs after Shield-1 removal, which provides an ideal opportunity to study the m A function with minimal steric interference from bulky dCas13b fusion protein.

摘要

N6-甲基腺苷(m A)在 RNA 上发挥着重要作用,调控着各种生物过程,CRIPSR 技术已被用于可编程 m A 编辑。然而,CRISPR 蛋白的庞大体积和组成型表达的 CRISPR/RNA 编辑酶会干扰靶 RNA 和细胞的固有功能。在此,我们报道了一种基于配体稳定的 dCas13 编辑器的条件性 m A 编辑平台(FKBP*-dCas13b-ALK)。通过添加或去除 Shield-1 分子,实现了该 m A 编辑系统的诱导表达。我们进一步证明,在 Shield-1 的控制下,靶向募集 dCas13b-m A 擦除融合蛋白和靶向 m A 擦除得以实现。此外,dCas13b 融合蛋白的释放和降解比 Shield-1 去除后靶 RNA 上 m A 的恢复更快,这为研究 m A 功能提供了一个理想的机会,可最大限度地减少庞大的 dCas13b 融合蛋白的空间干扰。

相似文献

5
8
Small Molecule-Inducible and Photoactivatable Cellular RNA N1-Methyladenosine Editing.小分子诱导和光活化的细胞 RNA N1-甲基腺苷编辑。
Angew Chem Int Ed Engl. 2024 Jun 21;63(26):e202320029. doi: 10.1002/anie.202320029. Epub 2024 May 16.

本文引用的文献

2
On demand CRISPR-mediated RNA N6-methyladenosine editing.按需进行的CRISPR介导的RNA N6-甲基腺苷编辑。
Genes Dis. 2022 Jun 3;9(6):1389-1390. doi: 10.1016/j.gendis.2022.05.019. eCollection 2022 Nov.
4
N-methyladenosine in poly(A) tails stabilize VSG transcripts.聚(A)尾中的 N6-甲基腺苷稳定 VSG 转录本。
Nature. 2022 Apr;604(7905):362-370. doi: 10.1038/s41586-022-04544-0. Epub 2022 Mar 30.
10
TRADES: Targeted RNA Demethylation by SunTag System.TRADES:利用SunTag系统进行靶向RNA去甲基化
Adv Sci (Weinh). 2020 Aug 16;7(19):2001402. doi: 10.1002/advs.202001402. eCollection 2020 Oct.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验