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Related Experiment Video

Updated: Mar 27, 2026

Generation of Cationic Nanoliposomes for the Efficient Delivery of In Vitro Transcribed Messenger RNA
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Cationic mRNA Lipid Nanoparticles for Ex Vivo NanoCAR-T Cell Engineering.

Laure Harinck1,2, Stijn De Munter2,3, Margo De Velder1,2

  • 1Laboratory for General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|March 24, 2026
PubMed
Summary

Cationic lipid nanoparticles (LNPs) enhance mRNA expression for CAR-T cell engineering, independent of cell activation or media complexity. This simplifies manufacturing and broadens therapeutic applications.

Keywords:
CAR‐T cellsimmunotherapylipid nanoparticlesmRNA deliverynon‐viral gene delivery

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Area of Science:

  • Biotechnology
  • Immunotherapy
  • Cellular Engineering

Background:

  • Lipid nanoparticles (LNPs) are promising non-viral vectors for mRNA delivery in CAR-T cell engineering.
  • The influence of LNP charge and transfection medium composition on T cell engineering requires further investigation.

Purpose of the Study:

  • To compare the efficacy of charge-neutral and cationic LNPs for ex vivo mRNA-based CAR-T cell engineering.
  • To evaluate the impact of LNP charge and medium composition on T cell transfection efficiency and viability.

Main Methods:

  • Comparison of charge-neutral C12-200 LNPs and cationic DOTAP(C12-200) LNPs for primary human T cell transfection.
  • Assessment of mRNA expression, cell viability, and cytotoxicity.
  • Analysis of LNP uptake mechanisms, including low-density lipoprotein receptor expression.

Main Results:

  • Charge-neutral LNPs showed efficient transfection and high viability in serum-free medium with apolipoprotein E, correlating with LDL receptor expression.
  • Cationic LNPs achieved superior mRNA expression regardless of medium or T cell activation but with lower cytocompatibility.
  • Both LNP types successfully delivered anti-CD20 nanoCAR mRNA, resulting in potent in vitro T cell cytotoxicity.

Conclusions:

  • LNP charge, medium composition, and T cell activation are critical factors in ex vivo T cell engineering.
  • Cationic LNPs enable robust, charge-dependent transfection, simplifying CAR-T cell manufacturing and potentially extending expression kinetics.
  • Cationic LNPs facilitate the engineering of less-activated T cells and offer a simplified approach to CAR-T cell production.