Preprint / Version 1

Global Incorporation of Synthetic ATP Analogs Reveals Poly(A)-Dependent Translation Differences in mRNA

This article is a preprint and has not been certified by peer review.

Authors

    Lu Zhou,  
    Lu Zhou
    • 中科院深圳先进技术研究院
    • Shenzhen Institutes of Advanced Technology image/svg+xml
    • 澳门大学
    Mengting Li,  
    Mengting Li
    Junlin Wen,  
    Junlin Wen
    Chunlei Zhang,  
    Chunlei Zhang
    • Gingko Biotech Limited
    Chenghe Xiong,  
    Chenghe Xiong
    Wenjian Ma,  
    Wenjian Ma
    Ziwen Dai,  
    Ziwen Dai
    Xuanjun Zhang,  
    Xuanjun Zhang
    • 澳门大学
    Hui Mei
    Hui Mei
Categories
Chemical Biology
Keywords
mRNA; N6-acylation; Poly(A) tail

Abstract

Chemical modifications of nucleosides are essential for enhancing the efficacy of therapeutic mRNAs. While uridine analogs like N1-methylpseudouridine (m1Ψ) are well studied, adenine modifications remain underexplored, despite adenine’s abundance and exclusive role in the poly(A) tail of eukaryotic mRNA. Inspired by the translational benefits of N4-acetylcytidine (ac4C), we designed and synthesized a series of novel N6-acylated ATP analogs. we systematically evaluated these analogs against established modifications, including N6-methyl (m6A), 2-amino (am2A) and 7-deaza (c7A) derivatives, by incorporating them into mRNAs via global substitution. Our investigation highlights the efficacy of the newly developed N6-acetyl (ac6A) modification. In non-polyadenylated mRNAs, ac6A substitution significantly enhanced translation, achieving a threefold increase over unmodified mRNA. Importantly, in polyadenylated mRNAs, ac6A mRNA maintained translation efficiency comparable to natural mRNA, demonstrating its high biocompatibility. Surprisingly, am2A modification displayed striking poly(A)-dependent translational behavior. While am2A modified mRNA showed a threefold translation increase in the absence of a poly(A) tail, its efficiency dropped to 6% of natural levels upon poly(A) addition. Structural simulations revealed that am2A group introduces steric clashed and electrostatic repulsion with poly(A) binding protein (PABP), hindering closed-loop formation and reducing translation. Overall, this work expands the chemical space of adenine modifications with effective N6-acylated analogs and highlights that region-specific modifications can be strategically exploited to optimize translation efficiency for mRNA therapeutic design.

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DOI:

https://doi.org/10.65215/LTSpreprints.2026.03.16.000160

Submission ID:

160

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Posted

2026-03-16

How to Cite

Zhou, L., Li, M., Wen, J., Zhang, C., Xiong, C., Ma, W., Dai, Z., Zhang, X., & Mei, H. (2026). Global Incorporation of Synthetic ATP Analogs Reveals Poly(A)-Dependent Translation Differences in mRNA. LangTaoSha Preprint Server. https://doi.org/10.65215/LTSpreprints.2026.03.16.000160

Declaration of Competing Interests

The authors declare no competing interests to disclose.

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The copyright holder for this preprint is the author/funder.

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This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.