5 years ago

Oligonucleotide-Addressed Covalent 3′-Terminal Derivatization of Small RNA Strands for Enrichment and Visualization

Oligonucleotide-Addressed Covalent 3′-Terminal Derivatization of Small RNA Strands for Enrichment and Visualization
Aleksandr Osipenko, Alexandra Plotnikova, Saulius Klimašauskas, Viktoras Masevičius, Giedrius Vilkaitis, Milda Nainytė
The HEN1 RNA 2′-O-methyltransferase plays important roles in the biogenesis of small non-coding RNAs in plants and proved a valuable tool for selective transfer of functional groups from cofactor analogues onto miRNA and siRNA duplexes in vitro. Herein, we demonstrate the versatile HEN1-mediated methylation and alkylation of small RNA strands in heteroduplexes with a range of complementary synthetic DNA oligonucleotides carrying user-defined moieties such as internal or 3′-terminal extensions or chemical reporter groups. The observed DNA-guided covalent functionalization of RNA broadens our understanding of the substrate specificity of HEN1 and paves the way for the development of novel chemo-enzymatic tools with potential applications in miRNomics, synthetic biology, and nanomedicine. Versatile RNA tagging: A method for addressable chemo-enzymatic labeling of miRNAs and siRNAs, which exploits the activity of HEN1 2′-O-methyltransferase to transfer functional groups to the 3′-end of the RNA strand in DNA/RNA heteroduplexes, is presented. This method permits a variety of user-defined single- or dual-reporter applications, in which one tag can be synthetically incorporated into the DNA probe and the second is covalently attached by HEN1 to the target RNA.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/anie.201701448

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