Abstract
We present a chemo-enzymatic approach for site-specific labeling of 5'-capped RNAs based on bioorthogonal chemistry. A trimethylguanosine synthase was engineered to transfer a terminal azido moiety to the 5'-cap which could be further modified using strain-promoted azide-alkyne cycloaddition (SPAAC).
Highlights
We present a chemo-enzymatic approach for site-specific labeling of 50-capped RNAs based on bioorthogonal chemistry
Some complex phenomena like the trafficking of mRNA which leads to subcellular mRNA localization should ideally be studied in living cells
Approaches based on chemical synthesis and metabolic labeling yielded numerous improvements during the last few years
Summary
We present a chemo-enzymatic approach for site-specific labeling of 50-capped RNAs based on bioorthogonal chemistry. An approach that (i) allows attachment of small fluorescent reporters to mRNA, (ii) is compatible with cellular components and (iii) not harmful to living cells would be a significant advance, regarding studies on mRNA trafficking and localization.
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