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Updated: Jun 28, 2026
Novel RNA-Binding Proteins Isolation by the RaPID Methodology
Published on: September 30, 2016
Isolation of a sequence-specific RNA binding protein, polypyrimidine tract binding protein, using RNA affinity
1Howard Hughes Medical Institute, University of California at Los Angeles, Los Angeles, California, USA.
Abstract:
Many important cellular processes are mediated by sequence-specific RNA binding proteins, and it is often necessary to purify these proteins. When the RNA binding site is known, it is convenient to use this RNA as a matrix for affinity purification. The intronic splicing silencer (ISS) element present upstream of the N1 exon of the c-src pre-mRNA is a high-affinity binding site for the polypyrimidine tract binding protein (PTB). Using a 5'-biotinylated ISS RNA and PTB as an example, I describe a one-step method for affinity chromatography of RNA binding proteins from nuclear extracts.
