ARiBo pull-down for riboproteomic studies based on label-free quantitative mass spectrometry.

ABSTRACT: As part of their normal life cycle, most RNA molecules associate with several proteins that direct their fate and regulate their function. Here, we describe a novel method for identifying proteins that associate with a target RNA. The procedure is based on the ARiBo method for affinity purification of RNA, which was originally developed to quickly purify RNA with high yields and purity under native conditions. The ARiBo method was further optimized using in vitro transcribed RNA to capture RNA-associating proteins from cellular extracts with high yields and low background protein contamination. For these RNA pull-downs, stem-loops present in the immature forms of let-7 miRNAs (miRNA stem-loops) were used as the target RNAs. Label-free quantitative mass spectrometry analysis allowed for the

SUBMITTER: Di Tomasso G 

PROVIDER: S-EPMC5066628 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature