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

Accepted version
Peer-reviewed

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Type

Article

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Authors

Schmidt, Karyn 
Buquicchio, Frank 
Carroll, Johanna S 
Distel, Robert J 
Novina, Carl D 

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.

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Keywords

MS2 coat protein, RNA immunoprecipitation, long non-coding RNA, ribonucleoprotein complex, scaffold, transcription factor

Journal Title

J Biol Methods

Conference Name

Journal ISSN

2326-9901
2326-9901

Volume Title

4

Publisher

Journal of Biological Methods

Rights

All rights reserved
Sponsorship
Cancer Research UK (C14303/A17197)