Electro-mechanical transfection for non-viral primary immune cell engineering

Non-viral approaches to transfection have emerged a viable option for gene transfer. Electro-mechanical transfection involving use of electric fields coupled with high fluid flow rates is a scalable strategy for cell therapy development and manufacturing. Unlike purely electric field-based or mechanical-based delivery methods, the combined effects result in delivery of genetic material at high efficiencies and low toxicity. This study focuses on delivery of reporter mRNA to show electro-mechanical transfection can be used successfully in human T cells. Rapid optimization of delivery to T cells was observed with efficiency over 90% and viability over 80%. Confirmation of optimized electro-mechanical transfection parameters was assessed in multiple use cases including a 50-fold scale up demonstration. Transcriptome and ontology analysis show that delivery, via electro-mechanical transfection, does not result in gene dysregulation. This study demonstrates that non-viral electro-mechanical transfection is an efficient and scalable method for cell and gene therapy engineering and development. One Sentence Summary This study demonstrates that non-viral electro-mechanical transfection is an efficient and scalable method for development of engineered cellular therapies. ### Competing Interest Statement CB and PG are founders and employees of Kytopen Corp. JS, JH, RM, RB, and BG were employees of Kytopen Corp at the time of manuscript creation. CB is a tenured professor and researcher at Massachusetts Institute of Technology.