RATA: A method for high-throughput identification of RNA bound transcription factors.

J Biol Methods
Authors
Abstract

Long non-coding RNAs (lncRNAs) regulate critical cellular processes and their dysregulation contributes to multiple diseases. Although only a few lncRNAs have defined mechanisms, many of these characterized lncRNAs interact with transcription factors to regulate gene expression, suggesting a common mechanism of action. Identifying RNA-bound transcription factors is especially challenging due to inefficient RNA immunoprecipitation and low abundance of many transcription factors. Here we describe a highly sensitive, user-friendly, and inexpensive technique called RATA (RNA-associated transcription factor array), which utilizes a MS2-aptamer pulldown strategy coupled with transcription factor activation arrays for identification of transcription factors associated with a nuclear RNA of interest. RATA requires only ~5 million cells and standard molecular biology reagents for multiplexed identification of up to 96 transcription factors in 2-3 d. Thus, RATA offers significant advantages over other technologies for analysis of RNA-transcription factor interactions.

Year of Publication
2017
Journal
J Biol Methods
Volume
4
Issue
1
Date Published
2017
ISSN
2326-9901
DOI
10.14440/jbm.2017.171
PubMed ID
29201933
PubMed Central ID
PMC5710740
Links
Grant list
R01 CA140986 / CA / NCI NIH HHS / United States
R01 CA185151 / CA / NCI NIH HHS / United States
T32 AI007386 / AI / NIAID NIH HHS / United States