Feng Chao, Chan Dalen, Spitale Robert C
Department of Pharmaceutical Sciences, University of California, Irvine, Irvine, CA, 92697, USA.
Department of Chemistry, University of California, Irvine, Irvine, CA, 92697, USA.
Methods Mol Biol. 2017;1648:247-256. doi: 10.1007/978-1-4939-7204-3_18.
RNA molecules have emerged as key players in nearly every facet of gene regulation. Such functions are governed by RNA's unique ability to fold into intricate secondary and tertiary structures. In order to understand, on the molecular level, how such structures from several chemical and enzymatic methods have been developed. One such method is RNA Selective Hydroxyl Acylation analyzed by Primer Extension, or SHAPE. SHAPE has emerged as a premier method of measuring RNA structure, with recent developments extending SHAPE into living cells. Here, we describe the use of SHAPE for measuring RNA structure inside living cells.
RNA分子已成为基因调控几乎各个方面的关键参与者。这些功能由RNA折叠成复杂二级和三级结构的独特能力所决定。为了在分子水平上理解这些结构是如何形成的,已经开发了几种化学和酶促方法。其中一种方法是通过引物延伸分析的RNA选择性羟基酰化,即SHAPE。SHAPE已成为测量RNA结构的主要方法,最近的进展将SHAPE扩展到活细胞中。在这里,我们描述了使用SHAPE测量活细胞内RNA结构的方法。