Optimizing Transposon-Based Gene Delivery For Cell Therapy Applications With The MaxCyte® ExPERT Scalable Electroporation Platform

Transposons are effective for gene transfer in cell engineering, offering advantages over viral vectors due to their cost-effectiveness and simpler manufacturing. This study explores optimizing the delivery of the piggyBac (PB) transposon system for engineering cell lines and primary T cells. Researchers aimed to maximize transposition efficiency, cell expansion, recovery, and functionality while limiting vector copy number.

Key findings indicate that GFP transfection efficiency and vector copy number (VCN) increase with transposon concentration, while expression durability improves with transposase concentration in K562 cells. For CD19 CAR-T cells, both transfection efficiency and VCN increased with transposon concentration, and cell yield generally improved with higher transposon-to-transposase ratios. An optimal 4:1 ratio balanced transposition efficiency, expression durability, and expansion, leading to consistent CD19 CAR expression, viability, yield, functional killing, phenotype, and VCN across multiple donors.

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