在体探测 RNA 结构。
Probing RNA structure in vivo.
机构信息
Department of Chemistry, The Pennsylvania State University, University Park, PA 16802, USA; Center for RNA Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA.
Center for RNA Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA; Department of Biology, The Pennsylvania State University, University Park, PA 16802, USA.
出版信息
Curr Opin Struct Biol. 2019 Dec;59:151-158. doi: 10.1016/j.sbi.2019.07.008. Epub 2019 Sep 13.
RNA structure underpins many essential functions in biology. New chemical reagents and techniques for probing RNA structure in living cells have emerged in recent years. High-throughput, genome-wide techniques such as Structure-seq2 and DMS-MaPseq exploit nucleobase modification by dimethylsulfate (DMS) to obtain complete structuromes, and are applicable to multiple domains of life and conditions. New reagents such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), glyoxal, and nicotinoyl azide (NAz) greatly expand the capabilities of nucleobase probing in cells. Additionally, ribose-targeting reagents in selective 2'-hydroxyl acylation and primer extension (SHAPE) detect RNA flexibility in vivo. These techniques, coupled with crosslinking nucleobases in psoralen analysis of RNA interactions and structures (PARIS), provide new and diverse ways to elucidate RNA secondary and tertiary structure in vivo and genome-wide.
RNA 结构是生物学中许多基本功能的基础。近年来，出现了用于探测活细胞中 RNA 结构的新的化学试剂和技术。高通量、全基因组技术，如 Structure-seq2 和 DMS-MaPseq，利用二甲硫酸盐（DMS）修饰核苷酸碱基来获得完整的结构组，并适用于多个生命领域和条件。新的试剂，如 1-乙基-3-(3-二甲基氨基丙基)碳二亚胺（EDC）、乙二醛和烟酰基叠氮化物（NAz），极大地扩展了细胞中核苷酸碱基探测的能力。此外，核糖靶向试剂在选择性 2'-羟基酰化和引物延伸（SHAPE）中检测体内 RNA 的灵活性。这些技术，加上交联 psoralen 分析 RNA 相互作用和结构（PARIS）中的核苷酸碱基，提供了新的和多样化的方法来阐明体内和全基因组范围的 RNA 二级和三级结构。
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