Nanopore microscope identifies RNA isoforms with structural colours
Nanopore microscope identifies RNA isoforms with structural colours
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DOI：
10.1038/s41557-022-01037-5
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发表时间：
2022-09-19
期刊：
影响因子：
21.8
通讯作者：
Keyser, Ulrich Felix
中科院分区：
文献类型：
--
作者：
Boskovic, Filip;Keyser, Ulrich Felix
识别RNA转录异构体需要复杂的协议，遭受各种酶的偏见。在这里，我们设计了三维分子结构，使识别转录异构体在单分子水平上使用固态纳米孔显微镜。我们用设计好的互补DNA链将靶RNA重折叠成RNA标识符。每个重塑的分子都带有一个独特的结构（伪）颜色序列。结构颜色由DNA结构、蛋白质标记、天然RNA结构或三者的组合组成。RNA标识符的结构颜色序列使得能够在没有预先扩增的情况下同时鉴定和相对定量多个RNA靶标。我们的无扩增RNA TargEt多重异构体传感（ARTEMIS）方法揭示了与已发表的变体一致的天然转录本的结构排列。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环状和线性转录异构体。
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.