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

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.

Keywords: Antibody; Native RNA immunoprecipitation; RNA extraction; RNA-binding protein; Real-time PCR.