Efficient recovery of the RNA-bound proteome and protein-bound transcriptome using phase separation (OOPS).

ABSTRACT: RNA-protein interactions play a pivotal role in cell homeostasis and disease, but current approaches to study them require a considerable amount of starting material, favor the recovery of only a subset of RNA species or are complex and time-consuming. We recently developed orthogonal organic phase separation (OOPS): a quick, efficient and reproducible method to purify cross-linked RNA-protein adducts in an unbiased way. OOPS avoids molecular tagging or the capture of polyadenylated RNA. Instead, it is based on sampling the interface of a standard TRIzol extraction to enrich RNA-binding proteins (RBPs) and their cognate bound RNA. OOPS specificity is achieved by digesting the enriched interfaces with RNases or proteases to release the RBPs or protein-bound RNA, respectively. Here we presen

SUBMITTER: Villanueva E 

PROVIDER: S-EPMC7613161 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature