Massively Parallel Dissection of RNA in RNA-protein Interactions in vivo

ABSTRACT: Many of the biological functions performed by RNA are mediated by RNA-binding proteins (RBPs), and understanding the molecular basis of these interactions is fundamental to molecular biology. Here, we present MPRNA-immunoprecipitation (MPRNA-IP), an adaptation of the previously developed massively parallel RNA assay (MPRNA) for in vivo high-throughput dissection of RNA-protein interactions, and describe statistical models for identifying RNA domains and parsing the structural contributions of RNA. By using custom pools of tens of thousands of RNA sequences containing systematically designed truncations and mutations, we are able to identify RNA domains, sequences, and secondary structures necessary and sufficient for protein binding in a single experiment. We show that this approach is successful for multiple RNAs of interest including NORAD, MS2, and human telomerase RNA, and we use it to interrogate the RNA-binding preferences of the DNA-looping factor CTCF. By blending modern and classical approaches, MPRNA-IP provides a novel high-throughput way to elucidate RNA-based mechanisms behind RNA-protein interactions.

ORGANISM(S): Homo sapiens

PROVIDER: GSE221537 | GEO | 2024/04/15

REPOSITORIES: GEO