Live-cell mapping of organelle-associated RNAs via proximity biotinylation combined with protein-RNA crosslinking.

ABSTRACT: The spatial organization of RNA within cells is a crucial factor influencing a wide range of biological functions throughout all kingdoms of life. However, a general understanding of RNA localization has been hindered by a lack of simple, high-throughput methods for mapping the transcriptomes of subcellular compartments. Here, we develop such a method, termed APEX-RIP, which combines peroxidase-catalyzed, spatially restricted in situ protein biotinylation with RNA-protein chemical crosslinking. We demonstrate that, using a single protocol, APEX-RIP can isolate RNAs from a variety of subcellular compartments, including the mitochondrial matrix, nucleus, cytosol, and endoplasmic reticulum (ER), with specificity and sensitivity that rival or exceed those of conventional approaches. We further

SUBMITTER: Kaewsapsak P 

PROVIDER: S-EPMC5730372 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature