Technical Note: Efficient Transfection of Lymphocytes with mRNA Using jetMESSENGER^®

Overview

Optimizing gene-delivery strategies for precise targeting of lymphocytes presents a significant challenge in both fundamental studies of T cells, B cells, and natural killer (NK) cells, as well as in clinical immunotherapy research. T lymphocytes, which are notoriously difficult to transfect, often rely on electroporation for gene delivery. However, this method can lead to issues such as low efficiency and reduced cell viability. Chemical transfection reagents commonly used for mammalian cells also show limited efficacy in T cells because T cells are resistant to many commercially available chemical transfection reagents, such as Lipofectamine™ 2000.

Despite the potential cellular damage associated with electroporation, it remains the predominant method for transfecting B cells. Additionally, in vitro delivery strategies for CRISPR/Cas9 into B cells usually depend on viral-based methods, which have limited carrier capacity and can cause immunogenicity issues and potential insertional mutagenesis. Modifying NK cells using conventional methods like electroporation, lipofection, or viral methods is even more challenging than modifying T cells, as NK cells are much more resistant. This has prompted the evaluation of mRNA transfection as an alternative approach.

To address these challenges, improved non-viral gene delivery methods that minimize cellular toxicity are required for effective gene transfer into lymphocytes. This study aims to assess and optimize the transfection efficiency and cell viability performance of jetMESSENGER^® reagent, a chemical-based transfection reagent for delivering mRNA into T lymphocytes, B lymphocytes, and NK cells.

• Document type: Technical Note
• Page count: 8
• Read time: 14 minutes

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