SHAPE-Seq: High-Throughput RNA Structure Analysis.
SHAPE-Seq: High-Throughput RNA Structure Analysis.
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DOI：
10.1002/9780470559277.ch120019
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发表时间：
2012-12-01
影响因子：
--
通讯作者：
Lucks, Julius B
中科院分区：
文献类型：
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作者：
Mortimer, Stefanie A;Trapnell, Cole;Lucks, Julius B
RNA结构的知识对于理解RNA在生物学中的重要功能作用和RNA的工程控制生物系统至关重要。本文包含了一种通过引物延伸和测序（SHAPE-Seq）分析的选择性2 '-羟基酰化的方案，该方案通过结构依赖性化学探测和下一代测序技术的组合，在一个实验中实现了数百种RNA的结构表征。该方案适用于各种条件，是理解RNA生物学的重要工具。该方案包括用于研究的RNA混合物的设计和合成的方法，以及揭示混合物中RNA的结构信息的结构依赖性测序文库的构建和分析。该方法通常适用于在各种条件下研究RNA结构和体外相互作用，并允许以高通量方式快速表征RNA结构。Curr.方案4：275-297 © 2012，John Wiley & Sons，Inc.
Knowledge of RNA structure is critical to understanding both the important functional roles of RNA in biology and the engineering of RNA to control biological systems. This article contains a protocol for selective 2'-hydroxyl acylation analyzed by primer extension and sequencing (SHAPE-Seq) that, through a combination of structure-dependent chemical probing and next-generation sequencing technologies, achieves structural characterization of hundreds of RNAs in a single experiment. This protocol is applicable in a variety of conditions, and represents an important tool for understanding RNA biology. The protocol includes methods for the design and synthesis of RNA mixtures for study, and the construction and analysis of structure-dependent sequencing libraries that reveal structural information of the RNAs in the mixtures. The methods are generally applicable to studying RNA structure and interactions in vitro in a variety of conditions, and allows for the rapid characterization of RNA structures in a high-throughput manner. Curr. Protoc. Chem. Biol. 4:275-297 © 2012 by John Wiley & Sons, Inc.