RNA-Binding Protein Immunoprecipitation from Whole-Cell Extracts

Köster T, Staiger D (2014)
Methods in molecular biology (Clifton, N.J.) 1062: 679-695.

Journal Article | Published | English

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Abstract
RNA-based regulation is increasingly recognized as an important factor shaping the cellular transcriptome. RNA-binding proteins that interact with cis-regulatory motifs within pre-mRNAs determine the fate of their targets. Understanding posttranscriptional networks controlled by an RNA-binding protein requires the identification of its immediate in vivo targets. Here we describe RNA immunoprecipitation in Arabidopsis thaliana. Transgenic plants expressing an RNA-binding protein fused to green fluorescent protein are treated with formaldehyde to "trap" RNAs in complexes with their physiological protein partners. A whole-cell extract is subjected to immunoprecipitation with an antibody against the GFP tag. In parallel, a mock immunoprecipitation is performed using an unrelated antibody. Coprecipitated RNAs are eluted from the immunoprecipitate and identified via real-time PCR. Enrichment relative to immunoprecipitation from plants expressing GFP only and mock immunoprecipitation with an unrelated antibody indicates specific binding.
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Köster T, Staiger D. RNA-Binding Protein Immunoprecipitation from Whole-Cell Extracts. Methods in molecular biology (Clifton, N.J.). 2014;1062:679-695.
Köster, T., & Staiger, D. (2014). RNA-Binding Protein Immunoprecipitation from Whole-Cell Extracts. Methods in molecular biology (Clifton, N.J.), 1062, 679-695.
Köster, T., and Staiger, D. (2014). RNA-Binding Protein Immunoprecipitation from Whole-Cell Extracts. Methods in molecular biology (Clifton, N.J.) 1062, 679-695.
Köster, T., & Staiger, D., 2014. RNA-Binding Protein Immunoprecipitation from Whole-Cell Extracts. Methods in molecular biology (Clifton, N.J.), 1062, p 679-695.
T. Köster and D. Staiger, “RNA-Binding Protein Immunoprecipitation from Whole-Cell Extracts”, Methods in molecular biology (Clifton, N.J.), vol. 1062, 2014, pp. 679-695.
Köster, T., Staiger, D.: RNA-Binding Protein Immunoprecipitation from Whole-Cell Extracts. Methods in molecular biology (Clifton, N.J.). 1062, 679-695 (2014).
Köster, Tino, and Staiger, Dorothee. “RNA-Binding Protein Immunoprecipitation from Whole-Cell Extracts”. Methods in molecular biology (Clifton, N.J.) 1062 (2014): 679-695.
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Polypyrimidine tract-binding proteins of potato mediate tuberization through an interaction with StBEL5 RNA.
Cho SK, Sharma P, Butler NM, Kang IH, Shah S, Rao AG, Hannapel DJ., J. Exp. Bot. 66(21), 2015
PMID: 26283046
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PMID: 25104024

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