nRIP-seq: a technique to identify RNA targets of an RNA binding protein on a genome-wide scale

Methods Mol Biol. 2015:1206:97-106. doi: 10.1007/978-1-4939-1369-5_9.

Abstract

Native RNA immunoprecipitation (nRIP) coupled with high-throughput sequencing (nRIP-seq) is a powerful technique that allows transcriptome-wide identification of the entire subset of coding and noncoding RNAs associated with a particular protein. Since this technology is carried out in a native condition without cross-linking, nRIP-seq detects RNAs that bind a protein directly or indirectly through a larger RNA-protein complex. Here, we use the interaction between RNA and chromatin modifiers, Polycomb proteins, as an example to describe this method. Using nRIP-seq, we provide a snapshot of Ezh2, a Polycomb component, and RNA interaction in mouse embryonic stem cells.

MeSH terms

  • Animals
  • Embryonic Stem Cells / metabolism
  • Gene Library
  • High-Throughput Nucleotide Sequencing
  • Immunoprecipitation / methods*
  • Mice
  • Polycomb-Group Proteins / metabolism
  • Polymerase Chain Reaction / methods
  • RNA / analysis
  • RNA / metabolism*
  • RNA-Binding Proteins / metabolism*

Substances

  • Polycomb-Group Proteins
  • RNA-Binding Proteins
  • RNA