Issue 17, 2022

Stronger together for in-cell translation: natural and unnatural base modified mRNA

Abstract

The preparation of highly modified mRNAs and visualization of their cellular distribution are challenging. We report in-cell application of in vitro transcribed mRNA containing natural base modifications and site-specifically introduced artificial nucleotides. Click chemistry on mRNA allows visualization in cells with excellent signal intensities. While non-specific introduction of reporter groups often leads to loss in mRNA functionality, we combined the benefits from site-specificity in the 3′-UTR incorporated unnatural nucleotides with the improved translation efficiency of the natural base modifications Ψ and 5mC. A series of experiments is described to observe, quantify and verify mRNA functionality. This approach represents a new way to visualize mRNA delivery into cells and monitor its spread on a cellular level and translation efficiency. We observed increased protein expression from this twofold chemically modified, artificial mRNA counterbalancing a reduced transfection rate. This synergetic effect can be exploited as a powerful tool for future research on mRNA therapeutics.

Graphical abstract: Stronger together for in-cell translation: natural and unnatural base modified mRNA

Supplementary files

Article information

Article type
Edge Article
Submitted
02 feb. 2022
Accepted
01 mar. 2022
First published
03 mar. 2022
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2022,13, 4753-4761

Stronger together for in-cell translation: natural and unnatural base modified mRNA

L. Bornewasser, C. Domnick and S. Kath-Schorr, Chem. Sci., 2022, 13, 4753 DOI: 10.1039/D2SC00670G

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