JS
Johanna Schott
  • Post-Doc, University Heidelberg Mannheim
Research fields
  • Biochemistry, Cancer Biology, Cell Biology, Immunology, Molecular Biology
Assessing the in vitro Binding Affinity of Protein–RNA Interactions Using an RNA Pull-down Technique
Authors:  Anand Chopra, Feras Balbous and Kyle K. Biggar, date: 12/05/2022, view: 2322, Q&A: 0

RNA is a vital component of the cell and is involved in a diverse range of cellular processes through a variety of functions. However, many of these functions cannot be performed without interactions with proteins. There are currently several techniques used to study protein–RNA interactions, such as electrophoretic mobility shift assay, fluorescence anisotropy, and filter binding. RNA-pulldown is a technique that uses biotinylated RNA probes to capture protein–RNA complexes of interest. First, the RNA probe and a recombinant protein are incubated to allow the in vitro interaction to occur. The fraction of bound protein is then captured by a biotin pull-down using streptavidin-agarose beads, followed by elution and immunoblotting for the recombinant protein with a His-tag–reactive probe. Overall, this method does not require specialized equipment outside what is typically found in a modern molecular laboratory and easily facilitates the maintenance of an RNase-free environment.


Graphical abstract



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