CRISPR-Cas13-Based RNA-Interacting Protein Detection in Living Cells

Abstract

Over the past decade, sequencing technology has revealed that the genomes of many organisms are pervasively transcribed, resulting in the production of numerous unknown RNA species. Systematic characterization of interacting proteins of these RNAs can advance our understanding of the function of diverse transcribed RNAs in gene expression regulation inside living cells. Zhang et al. have recently developed a programmable RNA-interacting protein detection tool named CRUIS.(1) They employed a hybrid of RNA-targeting CRISPR-Cas13 (LwaCas13a) and the proximity-labeling technology PUP-IT to achieve specific RNA docking and labeling of nearby proteins in living cells (Figure 1). Subsequently, the labeled proteins are enriched and identified by mass spectrometry. This method has been validated in abundant common RNA species including lncRNA and mRNA.