瑞典斯德哥尔摩大学Marc R. Friedländer和Inna Biryukova课题组合作的最新研究使用agoTRIBE揭示了单细胞全转录组中microRNA（miRNA）靶点相互作用。2023年9月21日出版的《自然-生物技术》发表了这项成果。

研究人员通过将miRNA效应蛋白Argonaute2整合到ADAR2的RNA编辑区域来克服这些限制，从而实现在单个细胞中检测转录组范围内的miRNA靶点。miRNA将融合蛋白引导到其天然靶转录本，对它们进行A>I编辑，并可以通过单细胞RNA测序来检测编辑效率。研究表明agoTRIBE可鉴定功能性miRNA靶点，这些靶点在进化序列保守性上相一致。

在该方法的一个应用中，研究人员探索了单细胞中microRNA的相互作用，并确定了整个细胞周期中的实质性差异靶点。AgoTRIBE还能对转录组范围内RNA丰度进行测量，并能在单细胞水平上对复杂组织中的miRNA靶向进行反卷积。

研究人员表示，MicroRNA通过对其靶基因转录本进行选择从而对许多生物过程发挥基因调控作用。目前用于鉴定miRNA靶标的实验方法很费力，需要数百万个细胞。

附：英文原文

Title: Detection of transcriptome-wide microRNA–target interactions in single cells with agoTRIBE

Author: Sekar, Vaishnovi, Mrmol-Snchez, Emilio, Kalogeropoulos, Panagiotis, Stanicek, Laura, Sagredo, Eduardo A., Widmark, Albin, Doukoumopoulos, Evangelos, Bonath, Franziska, Biryukova, Inna, Friedlnder, Marc R.

Issue&Volume: 2023-09-21

Abstract: MicroRNAs (miRNAs) exert their gene regulatory effects on numerous biological processes based on their selection of target transcripts. Current experimental methods available to identify miRNA targets are laborious and require millions of cells. Here we have overcome these limitations by fusing the miRNA effector protein Argonaute2 to the RNA editing domain of ADAR2, allowing the detection of miRNA targets transcriptome-wide in single cells. miRNAs guide the fusion protein to their natural target transcripts, causing them to undergo A>I editing, which can be detected by sensitive single-cell RNA sequencing. We show that agoTRIBE identifies functional miRNA targets, which are supported by evolutionary sequence conservation. In one application of the method we study microRNA interactions in single cells and identify substantial differential targeting across the cell cycle. AgoTRIBE also provides transcriptome-wide measurements of RNA abundance and allows the deconvolution of miRNA targeting in complex tissues at the single-cell level.

DOI: 10.1038/s41587-023-01951-0

Source: https://www.nature.com/articles/s41587-023-01951-0