Diazirine-Containing RNA Photo-Cross-Linking Probes for Capturing microRNA Targets

Abstract

Here, we report the applicability of diazirine-containing RNA photo-cross-linking probes for the identification of microRNA (miRNA) targets. The RNA cross-linking probes were synthesized by substituting the RNA nucleobases with nucleoside analogues such as 1-<i>O</i>-[3-(3-trifluoromethyl-3<i>H</i>-diazirin-3-yl)]­benzyl-β-d-ribofuranose or 1-<i>O</i>-[4-(3-trifluoromethyl-3<i>H</i>-diazirin-3-yl)]­benzyl-β-d-ribofuranose that carry aryl trifluoromethyl diazirine moieties. The probes were successfully cross-linked with synthetic RNAs containing the four natural nucleosides on the opposite site of the nucleoside analogues. Furthermore, it was found that miRNAs containing these analogues were effective in regulating the expression of their target genes. Thus, RNAs containing the nucleoside analogues are promising candidates as photo-cross-linking probes to identify the target mRNAs of miRNAs