RNA-Protein Interactions: PAR-CLIP Technology

This lecture introduces the PAR-CLIP technology, focusing on how it perturbs and probes cellular RNA/DNA using chemical biology toolsets like BrdU, EdU, and 'Spinach' probes. It explains the biological aim of probing RNA-protein complexes and reading RNA sequences. The lecture covers the process of culture cells with 4SU, crosslinking, immunoprecipitation, and sequencing to identify RNA-binding sites. It discusses the advantages and limitations of PAR-CLIP, including its accuracy in mapping interactions and potential cytotoxic effects. Additionally, it explores alternative applications of PAR-CLIP in capturing mRNA interactomes and tracking cellular RNAs using RNA aptamers as GFP mimics.