Enhancing CRISPR Knock-In And Knock-Out With The 4D-Nucleofector® LV PRO

The CRISPR/Cas9 system is transforming genome engineering, but moving from lab bench research to clinical applications for gene therapies requires safe and efficient delivery of its components into target cells. Electroporation stands out as the gold standard non-viral method due to its safety, flexibility, and increasing use in clinical trials.

This application note demonstrates the reliable, robust, and efficient non-viral delivery of complex, clinically relevant CRISPR cargo using the next-generation 4D-Nucleofector® LV Unit PRO and its Nucleocuvette® Cartridges PRO. The data confirms an easy and scalable workflow, matching performance from small-scale (100 µL) to large-scale (up to 20 mL and 1 x 10 9 cells) for T cell knock-in (KI) and knock-out (KO). The system maintains consistent efficiency across various input volumes, cell sources (activated and resting T cells), and process parameters, making it a valuable tool for non-viral manufacturing of gene and cell therapy products.

Explore the detailed results on scalability and robustness.