Scalable microfluidics that can address the challenge of delivery of large transgenes to therapeutic cells

To accelerate the manufacturing innovation, the goal of this project is to validate a cell engineering technology that uses a microfluidic transfection for delivery of therapeutic transgenes to four cell types of importance to the pharma industry.

Project status

100% Completed

Industry Need

Cost of manufacturing genetically engineered cells for therapies is significant, with high process complexity, and variable cell product quality.

Solution

Validate a cell engineering technology that uses microfluidic transfection for delivery of therapeutic transgenes.

Outputs/Deliverables

Developed consumable transfection chip for T cells and iPSCs that was delivered to partners
Demonstrated first mechanoporation transfection of iPSCs with large CRISPR-correction plasmids
Demonstrated high efficiency T cell knockout editing using mechanoporation
Demonstrated clinical scale processing (>1B cells) of T cells

Impacts

Demonstrate high efficiency T cell knockout editing using mechanoporation

Demonstrate mechanoporation transfection of iPSCs with large CRISPR-correction plasmids

Demonstrate clinical scale processing (>1B cells) of T cells

Develop a non-viral, mechanical approach to transfection of gene editing cargo, including CRISPR/Cas9, DNA, and mRNA

Publications

Loo, J., Sicher, I., Goff, A., Kim, O., Clary, N., Alexeev, A., Sulchek, T., Zamarayeva, A., Han, S., & Calero-Garcia, M. (2021). Microfluidic transfection of mRNA into human primary lymphocytes and hematopoietic stem and progenitor cells using ultra-fast physical deformations. Scientific Reports, 11(1). https://doi.org/10.1038/s41598-021-00893-4

Stone, N. E., Voigt, A. P., Mullins, R. F., Sulchek, T., & Tucker, B. A. (2021). Microfluidic processing of stem cells for autologous cell replacement. Stem Cells Translational Medicine, 10(10), 1384-1393. https://doi.org/10.1002/sctm.21-0080