Current approaches for RNA-labelling to identify RNA-binding proteins
Patel, Trushar R.
Canadian Science Publishing
RNA is involved in all domains of life, playing critical roles in a host of gene expression processes, host-defense mechanisms, cell proliferation, and diseases. A critical component in many of these events is the ability for RNA to interact with proteins. Over the past few decades, our understanding of such RNA–protein interactions and their importance has driven the search and development of new techniques for the identification of RNA-binding proteins. In determining which proteins bind to the RNA of interest, it is often useful to use the approach where the RNA molecule is the “bait” and allow it to capture proteins from a lysate or other relevant solution. Here, we review a collection of methods for modifying RNA to capture RNA-binding proteins. These include small-molecule modification, the addition of aptamers, DNA-anchoring, and nucleotide substitution. With each, we provide examples of their application, as well as highlight their advantages and potential challenges.
Open access article. Creative Commons Attribution 4.0 International license (CC BY 4.0) applies
RNA-binding proteins , Biotin labelling , Digoxigenin labelling , Aptamer labelling , Affinity capture , Nucleotide-substitution method
Gemmill, D., D'souza, S., Meier-Stephenson, V., & Patel, T. R. (2020). Current approaches for RNA-labelling to identify RNA-binding proteins. Biochemistry and Cell Biology, 98(1), 31-41. https://doi.org/10.1139/bcb-2019-0041