美国芝加哥大学Dickinson, Bryan C.团队报道了邻近依赖性RNA标记的生物正交掩膜酰化剂。相关研究成果发表在2024年4月9日出版的《自然—化学》。

RNA定位是高度调控的，随着亚细胞组织驱动依赖于环境的细胞生理学。尽管使用高反应性自由基或卡宾的基于接近度的标记技术，为细胞内蛋白质组织的无偏定位提供了一种强大的方法，但无偏RNA定位的方法很少，而且相对来说不那么稳健。

该文中，研究人员开发了α-烷氧基硫醇和氯烯醇酯，它们在受控的酯揭掩膜时起到强效酰化剂的作用。研究人员将这些探针与生物正交酯酶的亚细胞定位表达配对，以建立RNA空间分析的平台，用于邻近标记和测序的生物正交酰化剂（BAP-seq）。

研究证明，通过在感兴趣的区域选择性地揭开烯醇探针的面纱，可以绘制RNA在膜结合和无膜细胞器中的分布图。控释酰化剂化学和相应的BAP-seq方法扩大了接近标记技术的范围，并提供了一种强大的方法来访问RNA的细胞组织。

附：英文原文

Title: Bioorthogonal masked acylating agents for proximity-dependent RNA labelling

Author: Pani, Shubhashree, Qiu, Tian, Kentala, Kaitlin, Azizi, Saara-Anne, Dickinson, Bryan C.

Issue&Volume: 2024-04-09

Abstract: RNA localization is highly regulated, with subcellular organization driving context-dependent cell physiology. Although proximity-based labelling technologies that use highly reactive radicals or carbenes provide a powerful method for unbiased mapping of protein organization within a cell, methods for unbiased RNA mapping are scarce and comparably less robust. Here we develop α-alkoxy thioenol and chloroenol esters that function as potent acylating agents upon controlled ester unmasking. We pair these probes with subcellular-localized expression of a bioorthogonal esterase to establish a platform for spatial analysis of RNA: bioorthogonal acylating agents for proximity labelling and sequencing (BAP-seq). We demonstrate that, by selectively unmasking the enol probe in a locale of interest, we can map RNA distribution in membrane-bound and membrane-less organelles. The controlled-release acylating agent chemistry and corresponding BAP-seq method expand the scope of proximity labelling technologies and provide a powerful approach to interrogate the cellular organization of RNAs.

DOI: 10.1038/s41557-024-01493-1

Source: https://www.nature.com/articles/s41557-024-01493-1