Influenza A virus derived NS1 enhances translation of HPLC purified mRNA and interferon adjuvanted mRNA vaccination

influenza-a-virus-derived-ns1-enhances-translation-of-hplc-purified-mrna-and-interferon-adjuvanted-mrna-vaccination
Influenza A virus derived NS1 enhances translation of HPLC purified mRNA and interferon adjuvanted mRNA vaccination

Scientific Reports , Article number:  (2026) Cite this article

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Abstract

Non-structural protein (NS1) derived from influenza A virus is a potent immune evasion protein capable of enhancing translation of nucleoside-modified mRNA via host gene expression inhibition (HGEI) effect. In this study, we show that the HGEI effect on NS1 transfected cells reduces de novo synthesis and polyadenylation of host RNA. Co-delivering NS1 mRNA also enhances translation of non-inflammatory nucleoside-modified and HPLC-purified mRNA in NIH3T3 cells. Co-delivering NS1 mRNA also enhances translation of nucleoside-modified mRNA in DCs by 100% based on GFP expression, but completely inhibits cytokine induced maturation of primary human dendritic cells (DCs). Co-delivering NS1 mRNA also significantly enhances in vivo translation of naked mRNA via intramuscular route with transgene expression persisting for 12 days. As a proof of concept to mimic natural viral infection, when mRNAs encoding NS1 and interferon alpha were co-delivered, translation inhibition of the reporter gene was rescued both in vitro and in vivo. Using ovalbumin as a model antigen, we show that humoral response in C57BL/6 mice was enhanced when naked mRNAs encoding ovalbumin, NS1 and IFN were delivered together via intramuscular route. These novel properties of NS1 have the potential to impact mRNA therapeutics in significant ways.

Data availability

Data generated or analyzed during this study are included in this published article. Other datasets used and/or analyzed during the current study can be made available from the corresponding author on reasonable request.

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Funding

This work was funded by the Young Individual Research Grant from the National Medical Research Council of Singapore (NMRC/OFYIRG/0028/2016).

Author information

Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, 4 Engineering Drive 4, Singapore, 117585, Singapore

    Kyle K. L. Phua, Yi Liu, Wei Zhang & Jas-Min Chin

  2. Department of Surgery, Duke University School of Medicine, Durham, NC, 27710, USA

    Smita K. Nair

  3. Department of Medicine, University of Pennsylvania, 52 Johnson Pavilion, Philadelphia, PA, 19104, USA

    Norbert Pardi

  4. Program in Cancer and Stem Cell Biology, Duke-NUS Medical School, 8 College Road, Singapore, 169857, Singapore

    Hong-Wen Tang

  5. Stemigen Therapeutics, Lorong 8 Toa Payoh, Singapore, 319261, Singapore

    Kyle K. L. Phua

Authors

  1. Kyle K. L. Phua
  2. Yi Liu
  3. Wei Zhang
  4. Jas-Min Chin
  5. Hong-Wen Tang
  6. Norbert Pardi
  7. Smita K. Nair

Contributions

K.K.L.Phua conceived designed the study, wrote and edited the paper. Y Liu, W Zhang and J Chin performed the experiments. S K Nair performed experiments on human dendritic cells. N Pardi performed HPLC purification, H. Tang edited the paper.

Corresponding author

Correspondence to Kyle K. L. Phua.

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Competing interests

The authors declare no competing interests.

Ethics

Primary human cells used in these experiments were isolated from blood obtained from healthy human volunteers following informed written consent using a protocol approved by the Duke University Institutional Review Board and carried out in accordance with relevant guidelines and regulations. Prior to conducting the research described in this paper, the experimenters were trained in “Responsible Care and Use of Laboratory Animals” as required by the National Advisory Committee for Laboratory Animal Research (NACLAR) in Singapore. The facilities at National University of Singapore (NUS) are fully accredited by the American Association for Accreditation of Laboratory Animal Care (AAALAC) and all animal experiments were conducted in accordance with relevant guidelines and regulations and based on protocol approved by NUS Institutional Animal Care and Use Committee (IACUC). All methods were conducted in accordance with ARRIVE guidelines.

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Phua, K.K.L., Liu, Y., Zhang, W. et al. Influenza A virus derived NS1 enhances translation of HPLC purified mRNA and interferon adjuvanted mRNA vaccination. Sci Rep (2026). https://doi.org/10.1038/s41598-026-35611-5

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  • DOI: https://doi.org/10.1038/s41598-026-35611-5

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