Investigation of in vitro delivery conditions for self-amplifying mRNA

Wenting Li; Yiming Wang; Yi Xin Huo; Yuan Lu

doi:10.1007/s00449-025-03215-4

Investigation of in vitro delivery conditions for self-amplifying mRNA

Wenting Li
, Yiming Wang
, Yi Xin Huo^*
, Yuan Lu^*

^*Corresponding author for this work

School of Life Science

Research output: Contribution to journal › Article › peer-review

Abstract

Self-amplifying mRNA (SAM) shows promise for vaccines and gene therapy because of its self-replicating ability. However, current studies lack sufficient information for systematic parameter optimization and differentiation from conventional non-replicating mRNA (NRM). Therefore, the transfection efficiency of NRM and SAM platforms was evaluated by comparing delivery vectors and optimizing parameters for the SAM protocol. SAM and NRM showed similar transfection preferences, but their efficiencies differed. Optimized SAM transfection parameters were then established, including dose and incubation time. In this study, an in vitro multi-parameter delivery system for SAM was constructed, providing valuable insights into SAM transfection and its distinction from regular mRNA. This study contributes an experimental basis for the rational screening of nucleic acid drug carriers and the establishment of SAM multi-parameter evaluation criteria, and also lays an important foundation for optimizing low-dose immunization strategies and their clinical application translation.

Original language	English
Pages (from-to)	1833-1844
Number of pages	12
Journal	Bioprocess and Biosystems Engineering
Volume	48
Issue number	11
DOIs	https://doi.org/10.1007/s00449-025-03215-4
Publication status	Published - Nov 2025

Keywords

Drug delivery
Non-replicating mRNA
Nucleic acid
Self-amplifying mRNA
Transfection

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10.1007/s00449-025-03215-4

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title = "Investigation of in vitro delivery conditions for self-amplifying mRNA",

abstract = "Self-amplifying mRNA (SAM) shows promise for vaccines and gene therapy because of its self-replicating ability. However, current studies lack sufficient information for systematic parameter optimization and differentiation from conventional non-replicating mRNA (NRM). Therefore, the transfection efficiency of NRM and SAM platforms was evaluated by comparing delivery vectors and optimizing parameters for the SAM protocol. SAM and NRM showed similar transfection preferences, but their efficiencies differed. Optimized SAM transfection parameters were then established, including dose and incubation time. In this study, an in vitro multi-parameter delivery system for SAM was constructed, providing valuable insights into SAM transfection and its distinction from regular mRNA. This study contributes an experimental basis for the rational screening of nucleic acid drug carriers and the establishment of SAM multi-parameter evaluation criteria, and also lays an important foundation for optimizing low-dose immunization strategies and their clinical application translation.",

keywords = "Drug delivery, Non-replicating mRNA, Nucleic acid, Self-amplifying mRNA, Transfection",

author = "Wenting Li and Yiming Wang and Huo, \{Yi Xin\} and Yuan Lu",

note = "Publisher Copyright: {\textcopyright} The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2025.",

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T1 - Investigation of in vitro delivery conditions for self-amplifying mRNA

AU - Li, Wenting

AU - Wang, Yiming

AU - Huo, Yi Xin

AU - Lu, Yuan

N1 - Publisher Copyright: © The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2025.

PY - 2025/11

Y1 - 2025/11

N2 - Self-amplifying mRNA (SAM) shows promise for vaccines and gene therapy because of its self-replicating ability. However, current studies lack sufficient information for systematic parameter optimization and differentiation from conventional non-replicating mRNA (NRM). Therefore, the transfection efficiency of NRM and SAM platforms was evaluated by comparing delivery vectors and optimizing parameters for the SAM protocol. SAM and NRM showed similar transfection preferences, but their efficiencies differed. Optimized SAM transfection parameters were then established, including dose and incubation time. In this study, an in vitro multi-parameter delivery system for SAM was constructed, providing valuable insights into SAM transfection and its distinction from regular mRNA. This study contributes an experimental basis for the rational screening of nucleic acid drug carriers and the establishment of SAM multi-parameter evaluation criteria, and also lays an important foundation for optimizing low-dose immunization strategies and their clinical application translation.

AB - Self-amplifying mRNA (SAM) shows promise for vaccines and gene therapy because of its self-replicating ability. However, current studies lack sufficient information for systematic parameter optimization and differentiation from conventional non-replicating mRNA (NRM). Therefore, the transfection efficiency of NRM and SAM platforms was evaluated by comparing delivery vectors and optimizing parameters for the SAM protocol. SAM and NRM showed similar transfection preferences, but their efficiencies differed. Optimized SAM transfection parameters were then established, including dose and incubation time. In this study, an in vitro multi-parameter delivery system for SAM was constructed, providing valuable insights into SAM transfection and its distinction from regular mRNA. This study contributes an experimental basis for the rational screening of nucleic acid drug carriers and the establishment of SAM multi-parameter evaluation criteria, and also lays an important foundation for optimizing low-dose immunization strategies and their clinical application translation.

KW - Drug delivery

KW - Non-replicating mRNA

KW - Nucleic acid

KW - Self-amplifying mRNA

KW - Transfection

UR - https://www.scopus.com/pages/publications/105012579202

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DO - 10.1007/s00449-025-03215-4

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EP - 1844

JO - Bioprocess and Biosystems Engineering

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IS - 11

ER -

Investigation of in vitro delivery conditions for self-amplifying mRNA

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Keywords

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