RIP: an mRNA localization technique

Methods Mol Biol. 2011;714:407-22. doi: 10.1007/978-1-61779-005-8_25.

Abstract

A detailed understanding of post-transcriptional gene expression is necessary to correlate the different elements involved in the many levels of RNA-protein interactions that are needed to coordinate the cellular biomolecular machinery. The profile of mRNA, a major component of this machinery, can be examined after isolation from specific RNA-binding proteins (RBPs). RIP-Chip or ribonomic profiling is a versatilein vivo technique that has been widely used to study post-transcriptional gene regulation and the localization of mRNA. Here we elaborately detail the methodology for mRNA isolation using RBP immunoprecipitation (RIP) as a primary approach. Specific antibodies are used to target RBPs, which are then used to capture the associated mRNA.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies, Immobilized / immunology
  • Antibodies, Immobilized / metabolism
  • Antibody Specificity
  • Cell Extracts
  • Computational Biology
  • HeLa Cells
  • Humans
  • Immunoprecipitation / methods*
  • K562 Cells
  • Magnetics
  • Microspheres
  • Nerve Tissue Proteins / metabolism
  • Oligonucleotide Array Sequence Analysis
  • RNA Transport
  • RNA, Messenger / analysis*
  • RNA, Messenger / isolation & purification
  • RNA, Messenger / metabolism
  • Ribonucleoproteins / immunology
  • Ribonucleoproteins / isolation & purification
  • Ribonucleoproteins / metabolism
  • Staphylococcal Protein A / metabolism

Substances

  • Antibodies, Immobilized
  • Cell Extracts
  • G-substrate
  • Nerve Tissue Proteins
  • RNA, Messenger
  • Ribonucleoproteins
  • Staphylococcal Protein A