Enhancing cap-independent translation of linear mRNA
Enhancing cap-independent translation of linear mRNA
While cap-dependent translation remains the primary focus in mRNA-based therapeutics, cap-independent translation holds promise for targeting diseases ranging from cancer to neurodegeneration. However, cap-independently translated mRNAs are unstable, produce less protein than capped mRNAs, and current methods for their improvement are imperfect. Here, we propose the use of in vitro transcription priming with azido-modified dinucleotide primer and post-transcriptional modification utilising click chemistry to improve the properties of cap-independently translated mRNAs. Our results demonstrate a significant enhancement in mRNA stability and protein output without eliciting immunogenicity. Moreover, we show how the mRNA 5′-end modification strategy can be used to investigate transfection and cap-independent translation processes in cells overcoming burdens associated with previous methods. Together, our findings support cap-independent translation as a viable alternative to the established cap-dependent process and provide tools for further exploration and enhancement of this modality.
Golojuch, Sebastian
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Largey, Brendan
3d7cfa7a-6a9b-4ed8-b620-be327c076db5
El-Sagheer, Afaf
05b8295a-64ad-4fdf-ad57-c34934a46c04
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
16 October 2025
Golojuch, Sebastian
f7805c1d-8834-48e2-8ad9-0662b34cd69f
Largey, Brendan
3d7cfa7a-6a9b-4ed8-b620-be327c076db5
El-Sagheer, Afaf
05b8295a-64ad-4fdf-ad57-c34934a46c04
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Golojuch, Sebastian, Largey, Brendan, El-Sagheer, Afaf and Brown, Tom
(2025)
Enhancing cap-independent translation of linear mRNA.
Nature Communications, 16 (1), [9205].
(doi:10.1038/s41467-025-64257-6).
Abstract
While cap-dependent translation remains the primary focus in mRNA-based therapeutics, cap-independent translation holds promise for targeting diseases ranging from cancer to neurodegeneration. However, cap-independently translated mRNAs are unstable, produce less protein than capped mRNAs, and current methods for their improvement are imperfect. Here, we propose the use of in vitro transcription priming with azido-modified dinucleotide primer and post-transcriptional modification utilising click chemistry to improve the properties of cap-independently translated mRNAs. Our results demonstrate a significant enhancement in mRNA stability and protein output without eliciting immunogenicity. Moreover, we show how the mRNA 5′-end modification strategy can be used to investigate transfection and cap-independent translation processes in cells overcoming burdens associated with previous methods. Together, our findings support cap-independent translation as a viable alternative to the established cap-dependent process and provide tools for further exploration and enhancement of this modality.
Text
s41467-025-64257-6
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Accepted/In Press date: 9 September 2025
Published date: 16 October 2025
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© The Author(s) 2025.
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Local EPrints ID: 506840
URI: http://eprints.soton.ac.uk/id/eprint/506840
ISSN: 2041-1723
PURE UUID: 795cebba-c288-43fa-9b43-77ef7d9f0b99
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Date deposited: 18 Nov 2025 18:21
Last modified: 19 Nov 2025 02:40
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Author:
Sebastian Golojuch
Author:
Brendan Largey
Author:
Afaf El-Sagheer
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