RATA: A method for high-throughput identification of RNA bound transcription factors.
| 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
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| Journal | J Biol Methods
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| Volume | 4
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| Issue | 1
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| Date Published | 2017
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| ISSN | 2326-9901
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| DOI | 10.14440/jbm.2017.171
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| PubMed ID | 29201933
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| PubMed Central ID | PMC5710740
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| 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
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