Reference: Koh YY and Wickens M (2014) Determining the RNA specificity and targets of RNA-binding proteins using a three-hybrid system. Methods Enzymol 539:163-81

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Abstract


The three-hybrid system can be used to identify RNA sequences that bind a specific protein by screening a hybrid RNA library with a protein-activation domain fusion as 'bait.' These screens complement biochemical techniques, for example, SELEX, co-immunoprecipitation, and cross-linking experiments (see UV crosslinking of interacting RNA and protein in cultured cells and PAR-CLIP (Photoactivatable Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation): a step-by-step protocol to the transcriptome-wide identification of binding sites of RNA-binding proteins).

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Journal Article
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Koh YY, Wickens M
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