Journal
MOLECULAR PHARMACEUTICS
Volume 15, Issue 3, Pages 737-742Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.molpharmaceut.7b00356
Keywords
modified mRNA; RNA labeling; tetrazine ligation; tRNA guanine transglycosylase; RNA therapeutics
Funding
- U.S. Army Research Office through the MURI [W911NF-13-1-0383]
- San Diego Fellowship
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Modified mRNA (mod-mRNA) has recently been widely studied as the form of RNA useful for therapeutic applications due to its high stability and lowered immune response. Herein, we extend the scope of the recently established RNA-TAG (transglycosylation at guanosine) methodology, a novel approach for genetically encoded site specific labeling of large mRNA transcripts, by employing mod-mRNA as substrate. As a proof of concept, we covalently attached a fluorescent probe to mCherry encoding mod-mRNA transcripts bearing 5-methylcytidine and/or pseudouridine substitutions with high labeling efficiencies. To provide a versatile labeling methodology with a wide range of possible applications, we employed a two-step strategy for functionalization of the mod-mRNA to highlight the therapeutic potential of this new methodology. We envision that this novel and facile labeling methodology of mod-RNA will have great potential in decorating both coding and noncoding therapeutic RNAs with a variety of diagnostic and functional moieties.
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