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Updated: Nov 30, 2025

Novel RNA-Binding Proteins Isolation by the RaPID Methodology
Published on: September 30, 2016
In Vivo Cross-Linking Followed by polyA Enrichment to Identify Yeast mRNA Binding Proteins
1Department of Chemistry and Biochemistry, Loyola Marymount University, Los Angeles, CA, USA. sarah.mitchell@lmu.edu.
Abstract:
mRNA binding proteins regulate gene expression by controlling the processing, localization, decay, and translation of messenger RNAs (mRNAs). To fully understand these mechanisms of posttranscriptional gene regulation, it is necessary to identify the complete set of mRNA binding proteins. In recent years, several assays have been developed to accomplish this goal in a wide variety of organisms. This work describes a method for the systematic identification of mRNA binding proteins in Saccharomyces cerevisiae. This method applies in vivo UV cross-linking, affinity pull-down of polyA(+) mRNAs, and analysis by mass spectrometry to identify proteins that directly bind to mRNAs.

