美国华盛顿大学David M. Shechner课题组，通过寡核苷酸介导的近距离相互作用组图谱对RNA微环境进行多组学表征。该项研究成果于2024年10月28日在线发表在《自然—方法学》杂志上。

研究人员介绍了寡核苷酸介导的近距离相互作用组图谱（O-MAP）方法，其可用于阐明目标RNA周围的生物分子，并保持其原生环境。O-MAP使用类似RNA原位荧光杂交的寡核苷酸探针，将近距离生物素化酶送到目标RNA，从而使邻近分子通过链霉亲和素下拉富集。

这种方法引发了异常精确的生物素化，可以轻松优化并移植到新的目标或样本类型。研究人员以非编码RNA 47S、7SK和Xist为模型，开发了O-MAP工作流程，用于发现RNA邻近的蛋白质、转录本和基因组位点，从而实现这些RNA亚细胞区室及其新的调控相互作用的多组学特征化。O-MAP不需要基因编辑，仅使用现成的组件，并且所需细胞数量比现有方法少几个数量级，使其对大多数实验室都可行。

据了解，RNA分子形成复杂的分子相互作用网络，这些网络对其功能和细胞架构至关重要。然而，这些相互作用网络在原位探测时非常困难。

附：英文原文

Title: Multiomic characterization of RNA microenvironments by oligonucleotide-mediated proximity-interactome mapping

Author: Tsue, Ashley F., Kania, Evan E., Lei, Diana Q., Fields, Rose, McGann, Christopher D., Marciniak, Daphne M., Hershberg, Elliot A., Deng, Xinxian, Kihiu, Maryanne, Ong, Shao-En, Disteche, Christine M., Kugel, Sita, Beliveau, Brian J., Schweppe, Devin K., Shechner, David M.

Issue&Volume: 2024-10-28

Abstract: RNA molecules form complex networks of molecular interactions that are central to their function and to cellular architecture. But these interaction networks are difficult to probe in situ. Here, we introduce Oligonucleotide-mediated proximity-interactome MAPping (O-MAP), a method for elucidating the biomolecules near an RNA of interest, within its native context. O-MAP uses RNA-fluorescence in situ hybridization-like oligonucleotide probes to deliver proximity-biotinylating enzymes to a target RNA in situ, enabling nearby molecules to be enriched by streptavidin pulldown. This induces exceptionally precise biotinylation that can be easily optimized and ported to new targets or sample types. Using the noncoding RNAs 47S, 7SK and Xist as models, we develop O-MAP workflows for discovering RNA-proximal proteins, transcripts and genomic loci, yielding a multiomic characterization of these RNAs’ subcellular compartments and new regulatory interactions. O-MAP requires no genetic manipulation, uses exclusively off-the-shelf parts and requires orders of magnitude fewer cells than established methods, making it accessible to most laboratories.

DOI: 10.1038/s41592-024-02457-6

Source: https://www.nature.com/articles/s41592-024-02457-6