Streamlining CRISPR-Cas9 Gene Editing With LipidBrick® Cell Ready

By Sharlene Amador, Aaron Hoover, Chesney Michels, Jeff Cram, Mike Paglia

Electroporation has long been the standard for non-viral CRISPR delivery in T-cell engineering, but its impact on cell viability and expansion limits its scalability. Sartorius’ LipidBrick® Cell Ready offers a lipid-based alternative that preserves cell health while achieving comparable gene editing efficiency. In this study, researchers used LipidBrick® to deliver Cas9 mRNA and sgRNA for TRAC knockout in activated T cells. Optimization of the mRNA-to-sgRNA ratio, specifically a 1:11 ratio, proved critical for maximizing editing efficiency. Interestingly, increasing the total RNA payload did not improve knockout rates, and in some cases, lower RNA amounts performed better.

Liposome volume also played a key role, with larger volumes enhancing knockout efficiency without compromising cell viability. Compared to electroporation, LipidBrick®-treated cells showed faster recovery and better expansion, resulting in a higher yield of viable, TCR-depleted cells. These findings underscore the importance of delivery method, payload composition, and reagent volume in achieving efficient and scalable gene editing.

For teams developing cell therapies, LipidBrick® Cell Ready presents a compelling alternative to electroporation, one that supports healthier cells and higher product yields.