英国剑桥大学Keyser Ulrich Felix团队报道了纳米孔显微镜识别RNA异构体的结构颜色。相关研究成果于2022年9月19日发表于国际顶尖学术期刊《自然—化学》。
鉴定RNA转录亚型需要复杂的协议,这些协议会受到各种酶偏差的影响。
该文中,研究人员设计了三维分子结构,能够使用固态纳米孔显微镜在单分子水平上识别转录亚型。使用设计的互补DNA链将目标RNA重新折叠成RNA标识符。每个重塑的分子都有一系列独特的结构(伪)颜色。结构颜色由DNA结构、蛋白质标签、天然RNA结构或三者的组合组成。RNA标识符的结构颜色序列可以同时识别和相对量化多个RNA目标,而无需事先扩增。
该无扩增RNA TargEt多重异构体传感(ARTEMIS)方法揭示了天然转录物的结构排列,与已发表的变体一致。ARTEMIS使用单分子读数,在复杂的人类转录组中,通过一步无酶反应区分圆形和线性转录亚型。
附:英文原文
Title: Nanopore microscope identifies RNA isoforms with structural colours
Author: Bokovi, Filip, Keyser, Ulrich Felix
Issue&Volume: 2022-09-19
Abstract: Identifying RNA transcript isoforms requires intricate protocols that suffer from various enzymatic biases. Here we design three-dimensional molecular constructs that enable identification of transcript isoforms at the single-molecule level using solid-state nanopore microscopy. We refold target RNA into RNA identifiers with designed sets of complementary DNA strands. Each reshaped molecule carries a unique sequence of structural (pseudo)colours. Structural colours consist of DNA structures, protein labels, native RNA structures or a combination of all three. The sequence of structural colours of RNA identifiers enables simultaneous identification and relative quantification of multiple RNA targets without prior amplification. Our Amplification-free RNA TargEt Multiplex Isoform Sensing (ARTEMIS) method reveals structural arrangements in native transcripts in agreement with published variants. ARTEMIS discriminates circular and linear transcript isoforms in a one-step, enzyme-free reaction in a complex human transcriptome using single-molecule read-out.
DOI: 10.1038/s41557-022-01037-5
Source: https://www.nature.com/articles/s41557-022-01037-5
Nature Chemistry:《自然—化学》,创刊于2009年。隶属于施普林格·自然出版集团,最新IF:21.687
官方网址:https://www.nature.com/nchem/
投稿链接:https://mts-nchem.nature.com/cgi-bin/main.plex