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Biochemistry

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JoVE Journal Biochemistry
Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution
 

Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution

Article DOI: 10.3791/54391-v • 10:53 min • January 16th, 2017
January 16th, 2017 •
*1, *2, 2, 3, 1,2
1Department of Veterinary & Biomedical Sciences, University of Minnesota, 2Department of Veterinary Biosciences, Ohio State University, 3School of Biotechnology, Gautam Buddha University
* These authors contributed equally

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This manuscript describes an approach to isolate select cognate RNPs formed in eukaryotic cells via a specific oligonucleotide-directed enrichment. We demonstrate the applicability of this approach by isolating a cognate RNP bound to the retroviral 5' untranslated region that is composed of DHX9/RNA helicase A.

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Isolation RNA-protein Complexes Cells Oligonucleotide-directed Elution RNA Immunoprecipitation Technique Endogenous RNA Protein Complexes Eukaryotic Cells Retroviral Gene Expression Post-transcriptional Control Field RNA Binding Proteins Five Prime UTR Specific RNA Protein Complex Global Analysis RNP Downstream Viral Infections Cellular Diseases Mechanisms Of Post-transcriptional Control Drug Targets Culturing Cells Immunoprecipitation FLAG-tagged Protein Pro-TG Magnetic Bead Slurry Wash Buffer
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