Native RNA Immunoprecipitation (RIP) for Precise Detection and Quantification of Protein-Interacting RNA

Mai Baker; Rami Khosravi; Maayan Salton

doi:10.1007/978-1-0716-3191-1_8

Native RNA Immunoprecipitation (RIP) for Precise Detection and Quantification of Protein-Interacting RNA

Mai Baker, Rami Khosravi, Maayan Salton^*

^*Corresponding author for this work

Department of Biochemistry and Molecular Biology

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

1 Scopus citations

Abstract

Proteins with either RNA or DNA-binding motifs were shown to bind RNA. Immunoprecipitation of such proteins using antibodies and identification of the RNA-binding molecules is called RNA immunoprecipitation (RIP). The RNA precipitated with the studied protein can be detected by real-time polymerase chain reaction (PCR), microarray or sequencing. Here, we detail a method for native immunoprecipitation, without cross-linking, to isolate protein-RNA complexes followed by subsequent extraction and quantification of the co-purified RNA.

Original language	English
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	107-114
Number of pages	8
DOIs	https://doi.org/10.1007/978-1-0716-3191-1_8
State	Published - 2023

Publication series

Name	Methods in Molecular Biology
Volume	2666
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Publisher Copyright:
© 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

Antibody
Native RNA immunoprecipitation
RNA extraction
RNA-binding protein
Real-time PCR

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10.1007/978-1-0716-3191-1_8

Cite this

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doi = "10.1007/978-1-0716-3191-1_8",

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AB - Proteins with either RNA or DNA-binding motifs were shown to bind RNA. Immunoprecipitation of such proteins using antibodies and identification of the RNA-binding molecules is called RNA immunoprecipitation (RIP). The RNA precipitated with the studied protein can be detected by real-time polymerase chain reaction (PCR), microarray or sequencing. Here, we detail a method for native immunoprecipitation, without cross-linking, to isolate protein-RNA complexes followed by subsequent extraction and quantification of the co-purified RNA.

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KW - Real-time PCR

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Native RNA Immunoprecipitation (RIP) for Precise Detection and Quantification of Protein-Interacting RNA

Abstract

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Bibliographical note

Keywords

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