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Cited by 28RNA and protein synthesis mechanismsRNA Research and SplicingRNA modifications and cancer

Quantitative profiling of human translation initiation reveals elements that potently regulate endogenous and therapeutically modified mRNAs

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Lewis, Cole J.T., Xie, Li H., Bhandarkar, Shivani Milind, Jin, Danni, Abdallah, Kyrillos, Draycott, Austin S., Chen, Yixuan, Thoreen, Carson C. +1 more·2025·Molecular Cell
DOI 10.1016/j.molcel.2024.11.030PMC PMC11780321
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Abstract

mRNA therapeutics offer a potentially universal strategy for the efficient development and delivery of therapeutic proteins. Current mRNA vaccines include chemically modified nucleotides to reduce cellular immunogenicity. Here, we develop an efficient, high-throughput method to measure human translation initiation on therapeutically modified as well as endogenous RNAs. Using systems-level biochemistry, we quantify ribosome recruitment to tens of thousands of human 5' untranslated regions (UTRs) including alternative isoforms and identify sequences that mediate 200-fold effects. We observe widespread effects of coding sequences on translation initiation and identify small regulatory elements of 3-6 nucleotides that are sufficient to potently affect translational output. Incorporation of N1-methylpseudouridine (m1Ψ) selectively enhances translation by specific 5' UTRs that we demonstrate surpass those of current mRNA vaccines. Our approach is broadly applicable to dissecting mechanisms of human translation initiation and engineering more potent therapeutic mRNAs.

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Cited by

Sorted by citations

Authors
Lewis, C. J. T., Xie, L. H., Bhandarkar, S. M., Jin, D., Abdallah, K., Draycott, A. S., Chen, Y., Thoreen, C. C., & Gilbert, W. V.
Affiliations
Yale University (US)
Corresponding author
  • Thoreen, Carson C.
  • Gilbert, Wendy V.
ORCID
Venue
Molecular Cell
Publisher
Elsevier BV
Published
2025-01
Volume
85
Issue
2
Pages
445-459.e5
ISSN
1097-2765 (ISSN-L)
DOI
10.1016/j.molcel.2024.11.030
PMC
PMC11780321
OpenAlex
W4405567020
Cited by
28
References
3
Citations by year
2026: 13 · 2025: 13 · 2024: 2
FWCI
3.62
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Cell (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • SSRN Electronic Journal (acceptedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed Central (submittedVersion)
Funders
  • Pfizer (ITEN2021.RNA01)
  • National Cancer Institute (F31CA254339)
  • National Institute of Diabetes and Digestive and Kidney Diseases (F31DK129022)
  • National Institute of General Medical Sciences (R01GM132358)
  • National Science Foundation (2330451, DGE2139841)
  • American Cancer Society (PF-23-1144579-01-RMC)
  • National Institutes of Health (R35GM152167)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA modifications and cancer
Keywords
translation initiation, mRNA therapeutics, mRNA vaccines, chemically modified nucleotides, N1-methylpseudouridine, 5' untranslated region, ribosome recruitment

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