RNA-Binding Protein Immunoprecipitation from Whole-Cell Extracts

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

Download
Es wurde kein Volltext hochgeladen. Nur Publikationsnachweis!
Zeitschriftenaufsatz | Veröffentlicht | Englisch
Abstract / Bemerkung
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.
Erscheinungsjahr
Zeitschriftentitel
Methods in molecular biology (Clifton, N.J.)
Band
1062
Seite
679-695
ISSN
PUB-ID

Zitieren

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. doi:10.1007/978-1-62703-580-4_35
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.

9 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

PA5470 Counteracts Antimicrobial Effect of Azithromycin by Releasing Stalled Ribosome in Pseudomonas aeruginosa.
Shi J, Liu Y, Zhang Y, Jin Y, Bai F, Cheng Z, Jin S, Wu W., Antimicrob Agents Chemother 62(2), 2018
PMID: 29203495
E+ subgroup PPR protein defective kernel 36 is required for multiple mitochondrial transcripts editing and seed development in maize and Arabidopsis.
Wang G, Zhong M, Shuai B, Song J, Zhang J, Han L, Ling H, Tang Y, Wang G, Song R., New Phytol 214(4), 2017
PMID: 28277611
Adaptation of iCLIP to plants determines the binding landscape of the clock-regulated RNA-binding protein AtGRP7.
Meyer K, Köster T, Nolte C, Weinholdt C, Lewinski M, Grosse I, Staiger D., Genome Biol 18(1), 2017
PMID: 29084609
N(6)-Methyladenosine RNA Modification Regulates Shoot Stem Cell Fate in Arabidopsis.
Shen L, Liang Z, Gu X, Chen Y, Teo ZW, Hou X, Cai WM, Dedon PC, Liu L, Yu H., Dev Cell 38(2), 2016
PMID: 27396363
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
Regulation of pri-miRNA processing by the hnRNP-like protein AtGRP7 in Arabidopsis.
Köster T, Meyer K, Weinholdt C, Smith LM, Lummer M, Speth C, Grosse I, Weigel D, Staiger D., Nucleic Acids Res 42(15), 2014
PMID: 25104024

Export

Markieren/ Markierung löschen
Markierte Publikationen

Open Data PUB

Quellen

PMID: 24057392
PubMed | Europe PMC

Suchen in

Google Scholar