Click to Translate: Synthesis of Trans-Cyclooctene Modified 5' mRNA Caps for Bioorthogonal Activation
Niclas Zips1, Ekaterina Kulko1, Stephanie Kath-Schorr1
1Department of Chemistry and Biochemistry, University of Cologne, Köln, Germany.
Abstract:
mRNA translation can be controlled through chemical modification of the 5'-Cap, yet systematic insights into structure-reactivity relationships of bioorthogonally activatable Cap analogues remain limited. Here, we report the synthesis of trans-cyclooctene (TCO)-modified 5' mRNA caps (ZipCaps) and a comparative analysis of distinct TCO isomers. A controlled synthetic strategy enabled access to axial and equatorial derivatives, revealing stereochemical effects on click-to-release reactivity and stability. ZipCap-modified mRNA exhibits reduced translation that can be restored upon tetrazine addition, enabling reversible chemical control of protein expression in cells. In contrast to a recent report describing complete translational suppression, residual translation is observed, highlighting the sensitivity of Cap-dependent translation to structural variations.
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