Genome-wide mapping of RNA structure using nuclease digestion and high-throughput sequencing.

Document Type

Article

Publication Date

5-2013

Keywords

Gene Library, Genomics, High-Throughput Nucleotide Sequencing, Models, Molecular, Nucleic Acid Conformation, RNA, Fungal, Sequence Analysis, RNA, Yeasts

Volume

8

Issue

5

First Page

849

Last Page

869

ISSN

1750-2799

PMID

23558785

Abstract

RNA structure is important for RNA function and regulation, and there is growing interest in determining the RNA structure of many transcripts. Here we provide a detailed protocol for the parallel analysis of RNA structure (PARS) for probing RNA secondary structures genome-wide. In this method, enzymatic footprinting is coupled to high-throughput sequencing to provide secondary structure data for thousands of RNAs simultaneously. The entire experimental protocol takes ∼5 d to complete, and sequencing and data analysis take an additional 6-8 d. PARS was developed using the yeast genome as proof of principle, but its approach should be applicable to probing RNA structures from different transcriptomes and structural dynamics under diverse solution conditions. Nat Protoc 2013 May; 8(5):849-69.

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