Metabolic Incorporation of Azide Functionality into Cellular RNA.
Real-time tracking of RNA expression can provide insight into the mechanisms used to generate cellular diversity, as well as help determine the underlying causes of disease. Here we present the exploration of azide-modified nucleoside analogues and their ability to be metabolically incorporated into cellular RNA. We report robust incorporation of adenosine analogues bearing azide handles at both the 2'- and N6-positions; 5-methylazidouridine was not incorporated into cellular RNA. We further demonstrate selectivity of our adenosine analogues for transcription and polyadenylation. We predict that azidonucleosides will find widespread utility in examining RNA functions inside living cells, as well as in more complex systems such as tissues and living animals.
Department of Pharmaceutical Sciences, University of California, 2403 Natural Sciences I, Irvine, CA 92617, USA.
Sarah
S
Nainar
Department of Pharmaceutical Sciences, University of California, 2403 Natural Sciences I, Irvine, CA 92617, USA.
Samantha
S
Beasley
Department of Pharmaceutical Sciences, University of California, 2403 Natural Sciences I, Irvine, CA 92617, USA.
Michael
M
Fazio
Department of Pharmaceutical Sciences, University of California, 2403 Natural Sciences I, Irvine, CA 92617, USA.
Miles
M
Kubota
New England Biolabs, 240 County Road, Ipswich, MA 01938, USA.
Nan
N
Dai
New England Biolabs, 240 County Road, Ipswich, MA 01938, USA.
Ivan R
IR
Corrêa
Department of Pharmaceutical Sciences, University of California, 2403 Natural Sciences I, Irvine, CA 92617, USA.|Department of Chemistry, University of California, 2403 Natural Sciences I, Irvine, CA 92617, USA.
Robert C
RC
Spitale
D000241:Adenosine / Q000031:analogs & derivatives* / Q000378:metabolism; D000480:Alkynes / Q000737:chemistry; D001386:Azides / Q000737:chemistry*; D002384:Catalysis; D003300:Copper / Q000737:chemistry; D061565:Cycloaddition Reaction; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D009705:Nucleosides / Q000737:chemistry*; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D012264:Ribonucleotide Reductases / Q000037:antagonists & inhibitors
D000480:Alkynes; D001386:Azides; D005456:Fluorescent Dyes; D009705:Nucleosides; D012313:RNA; D003300:Copper; D012264:Ribonucleotide Reductases; D000241:Adenosine
RNA incorporation; azides; imaging; modified nucleosides; transcription