Temporal Labeling of Nascent RNA Using Photoclick Chemistry in Live Cells

Sarah Nainar; Miles Kubota; Christopher McNitt; Catherine Tran; Vladimir V Popik; Robert C Spitale

doi:10.1021/jacs.7b03121

Temporal Labeling of Nascent RNA Using Photoclick Chemistry in Live Cells

J Am Chem Soc. 2017 Jun 21;139(24):8090-8093. doi: 10.1021/jacs.7b03121. Epub 2017 Jun 8.

Authors

Sarah Nainar, Miles Kubota, Christopher McNitt¹, Catherine Tran, Vladimir V Popik¹, Robert C Spitale

Affiliation

¹ Department of Chemistry, University of Georgia , 140 Cedar Street, Athens, Georgia 30602, United States.

PMID: 28562039
DOI: 10.1021/jacs.7b03121

Abstract

We report the first cellular application of a photoclick SPAAC reagent to label azide-functionalized RNA. 350 nm irradiation of a cyclopropenone caged oxo-dibenzocyclooctyne (photo-ODIBO) biotin yields formation of the SPAAC reactive species, which rapidly forms adducts with RNA containing 2'-azidoadenosine (2'N₃-A). Photo-ODIBO was found to be highly stable in the presence of thiols, conferring greater stability relative to ODIBO. Light activated photo-ODIBO enabled tagging of cellular RNA, in addition to fluorescent imaging as well as enrichment of RNA in cell subpopulations via selective irradiation.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Alkynes / chemistry*
Cell Survival
Click Chemistry*
Cyclopropanes / chemistry*
HeLa Cells
Humans
Molecular Structure
Photochemical Processes*
RNA / analysis*
RNA / biosynthesis*
RNA / chemistry

Substances

Alkynes
Cyclopropanes
cyclopropenone
1-octyne
RNA