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Capture and Identification of miRNA Targets by Biotin Pulldown and RNA-seq. | LitMetric

Capture and Identification of miRNA Targets by Biotin Pulldown and RNA-seq.

Methods Mol Biol

Cellular and Molecular Medicine Program, Boston Children's Hospital, Boston, MA, 02115, USA.

Published: July 2016

AI Article Synopsis

Article Abstract

MicroRNAs (miRNAs) are small noncoding RNAs that regulate the stability and expression of target RNAs in a sequence-dependent manner. Identifying miRNA-regulated genes is key to understanding miRNA function. Here, we describe an unbiased biochemical pulldown method to identify with high-specificity miRNA targets. Regulated transcripts are enriched in streptavidin-captured mRNAs that bind to a transfected biotinylated miRNA mimic. The method is relatively simple, does not involve cross-linking and can be performed with only a million cells. Addition of an on-bead RNase digestion step also identifies miRNA recognition elements (MRE).

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Source
http://dx.doi.org/10.1007/978-1-4939-3067-8_13DOI Listing

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