Identification of sRNA interacting with a transcript of interest using MS2-affinity purification coupled with {RNA} sequencing (MAPS) technology

摘要

Abstract {RNA} sequencing (RNAseq) technology recently allowed the identification of thousands of small {RNAs} (sRNAs) within the prokaryotic kingdom. However, drawing the comprehensive interaction map of a sRNA remains a challenging task. To address this problem, we recently developed a method called {MAPS} (MS2 affinity purification coupled with {RNA} sequencing) to characterize the full targetome of specific sRNAs. This method enabled the identification of target {RNAs} interacting with sRNAs, regardless of the type of regulation (positive or negative), type of targets (mRNA, tRNA, sRNA) or their abundance. We also demonstrated that we can use this technology to perform a reverse {MAPS} experiment, where an {RNA} fragment of interest is used as bait to identify interacting sRNAs. Here, we demonstrated that RybB and MicF sRNAs co-purified with internal transcribed spacers (ITS) of metZ–metW–metV tRNA transcript, confirming results obtained with MS2-RybB MAPS. Both raw and analyzed {RNAseq} data are available in {GEO} database (GSE66517).