Vinyluridine as a Versatile Chemoselective Handle for the Post-transcriptional Chemical Functionalization of RNA

The development of modular and efficient methods to functionalize RNA with biophysical probes is very important in advancing the understanding of the structural and functional relevance of RNA in various cellular events. Herein, we demonstrate a two-step bioorthogonal chemical functionalization appr...

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Bibliographic Details
Published in:Bioconjugate chemistry 2017-05, Vol.28 (5), p.1529-1536
Main Authors: George, Jerrin Thomas, Srivatsan, Seergazhi G
Format: Article
Language:English
Subjects:
Acids
Animals
Cellular structure
Chemicals
Conjugation
Cycloaddition Reaction
DNA probes
Electrons
Humans
In vitro methods and tests
Nucleotide analogs
Palladium
Post-transcription
Probes
Ribonucleic acid
RNA
RNA - chemistry
RNA probes
RNA Processing, Post-Transcriptional
Structure-function relationships
Substrates
Transcription
Transcription factors
Uridine
Uridine Triphosphate - chemistry
Vinyl Compounds - chemistry
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Summary:The development of modular and efficient methods to functionalize RNA with biophysical probes is very important in advancing the understanding of the structural and functional relevance of RNA in various cellular events. Herein, we demonstrate a two-step bioorthogonal chemical functionalization approach for the conjugation of multiple probes onto RNA transcripts using a 5-vinyl-modified uridine nucleotide analog (VUTP). VUTP, containing a structurally noninvasive and versatile chemoselective handle, was efficiently incorporated into RNA transcripts by in vitro transcription reactions. Furthermore, we show for the first time the use of a palladium-mediated oxidative Heck reaction in functionalizing RNA with fluorogenic probes by reacting vinyl-labeled RNA transcripts with appropriate boronic acid substrates. The vinyl label also permitted the post-transcriptional functionalization of RNA by a reagent-free inverse electron demand Diels–Alder (IEDDA) reaction in the presence of tetrazine substrates. Collectively, our results demonstrate that the incorporation of VUTP provides newer possibilities for the modular functionalization of RNA with variety of reporters.
ISSN:1043-1802
1520-4812
DOI:10.1021/acs.bioconjchem.7b00169