Talk abstracts

Talk on Saturday 12:00-12:15pm submitted by Eduardo Paredes

Click Chemistry for RNA: Labeling and Ligation

Eduardo Paredes (Department of Chemistry, Carnegie Mellon University), Subha R. Das (Department of Chemistry, Carnegie Mellon University)

Abstract:
The Huisgen cycloaddition (Click Chemistry) involves a copper(I)-catalyzed triazole formation from an azide and an alkyne. The azide and alkyne moieties can be used interchangeably, one to tag the molecule of interest and the other in the molecule used for the detection, e.g. DNA and a fluorescent dye respectively. This reaction is not viable in biological systems due to the instability of the Cu(I) catalyst under oxygen rich aqueous solutions. Several stabilizing ligands have been reported that enable this reaction to be used in biological systems containing DNA. The use of systems containing RNA is specially challenging as the Cu(I) disproportionation reaction degrades RNA. Here we report the application of Click chemistry to RNA and a 5’end labeling of RNA with a fluorescent dye. This obviates the need to work with radioactively labeled RNA to study ribozyme kinetics and provides a sensitive and more stable probe that is readily accessible via Click Chemistry. In addition, We have synthesized a 3’-alkyne terminated RNA via solid phase synthesis and used 5’-deoxy 5’-azido guanosine according to literature to furnish a 5’-deoxy-5’azido transcript. Clicking these two fragments yields a ‘Click-Ligated’ RNA.

Keywords: Click Chemistry, RNA, Labeling