mRNA Electroporation Services
For therapeutic application, these methods are required to efficient transfection of many cells in a process with high cell survival. Among them, electroporation (EP) is a broadly applicable and a well-established method for transient mRNA transfection in a variety of different cell types. With years of optimizing and adapting, this method has been applied to yield high numbers of viable, transgene expressing dendritic cells (DCs), B cells, T cells, adult stem cells and several cell lines. The applications of IVT mRNA EP include cancer immunotherapy, protein replacement, and gene editing. For instance, EP is a commonly used method for preparing DC-based mRNA vaccines. Presently, several DC-based mRNA vaccines generated by electroporation have been evaluated in clinical trials. With state-of-the-art instruments and professional training of technicians, Creative Biogene is proud to offer mRNA EP service to satisfy our global customers' specific project needs. Our service is based on the EP method with a continuous flow system. Therefore, our service is flexible and scalable. We can support to transfect IVT mRNA into adherent and suspension cells with transfection rates of >75 % and survival rates of >90 %.
The advantages of mRNA electroporation
Electroporation (EP) is one of the oldest transfection methods, increasing eukaryotic cell membrane permeability to nucleic acids using an electrical field. Although the exact mechanism that how the membrane passed still needs to be explored, one thing is clear that nucleic acid moves in part because of its electric charge in the electric field. mRNA EP may be defined as genetic engineering. However, since there is no risk of integration into the host genome, mRNA has a large advantage over DNA. Here, we present several features and benefits of mRNA EP, including,
- Easy adaptation to different cell types
- High transfection efficiency and high reproducibility
- The rapid and transient expression
- The method possesses scalability and GMP compatibility property
- Homogenous protein expression
- No transgene integration into the genome
- Absence of an influence on cell phenotype
Fig1. The principle of electroporation. (Du, X., et al., 2018)
Service offering in Creative Biogene
To achieve IVT mRNA transfection into eukaryotic cells, we offer the EP method in terms of high gene transfection efficiency and low transfection-related toxicity. Since mRNA EP disrupted the cell membrane through an electric shock to enable intracellular mRNA delivery, we particularly focus on the adjustion of electrical characteristics, cell number, RNA quantity and other parameters, to assure the best optimized condition. The optimized EP conditions used in our service have shown no significant alteration of cells viability, proliferation and apoptosis induction, although many transfection methods may induce unforeseen adverse affects on transfected cells. In our service, we strive to maintain the biological properties of transfected cells in terms of their phenotypes, maturation status, and cytokine secreting ability, and among others. In addition, the availability of the continuous flow system makes our service amenable to high-throughput screening protocols, and able to large-scale genetic modification of cells required for therapeutic applications.
Electroporation conditions for different types of cells. (Gerer, K. F., et al., 2017)
In total, mRNA EP is a powerful tool to introduce genes easily into eukaryotic cells with high efficiency and low toxicity. Creative Biogene promises to provide high-quality and low-cost IVT mRNA transfection service tailored to customers' projects. If you are interested in our services, please contact us for more information. We are glad to cooperate with you!
References
- Du, X., et al. (2018). "Advanced physical techniques for gene delivery based on membrane perforation." Drug delivery, 25(1), 1516-1525.
- Gerer, K. F., et al. (2017). "Electroporation of mRNA as universal technology platform to transfect a variety of primary cells with antigens and functional proteins." RNA Vaccines. 2017. 165-178.