Project description:Goal was to identify RNA-binding proteins with a preference for m6A-modified RNA using an RNA bait generated from a known m6A site in the Dlg4 (PSD-95) mRNA 3`-UTR. By comparison of differential enrichment between m6A-RNA pulldown fractions and A-RNA or no RNA fractions, we can identify m6A-binding proteins.
Project description:Here we show that biotin-labelled miR-34a can be loaded to AGO2, and AGO2 immunoprecipitation can pulldown biotinylated miR-34a (Bio-miR pulldown). RNA-sequencing (RNA-seq) of the Bio-miR pulldown RNAs efficiently identified miR-34a mRNA targets, which could be verified with luciferase assays. In contrast to the approach of Bio-miR pulldown, RNA-seq of miR-34a overexpression samples had limited value in identifying direct targets of miR-34a. It seems that pulldown of 30 -Biotin-tagged miRNA can identify bona fide microRNA targets at least for miR34a.
Project description:Goal was to identify RNA-binding proteins with a preference for m6A-modified RNA using an RNA bait generated from a known m6A site in the Dlg4 (PSD-95) mRNA 3`-UTR. By comparison of differential enrichment between m6A-RNA pulldown fractions and A-RNA or no RNA fractions, we can identify m6A-binding proteins.
Project description:In order to find the protein specifically bound by lncRNA-AC016745.3, we identified it through RNA pulldown combined with Protein Mass Spectrometry