Temporal Labeling of Nascent RNA Using Photoclick Chemistry in Live Cells.

Sarah NainarMiles KubotaChristopher McNittCatherine TranVladimir V PopikRobert C Spitale

Published in: Journal of the American Chemical Society (2017)

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.

Keyphrases

nucleic acid
induced apoptosis
stem cells
photodynamic therapy
radiation therapy
bone marrow
oxidative stress
mass spectrometry
endoplasmic reticulum stress