Simple, fast and efficient iTOP-mediated delivery of CRISPR/Cas9 RNP in difficult-to-transduce human cells including primary T cells

The advent of the CRISPR/Cas9 system has transformed field human genome engineering and created new perspectives in development innovative cell therapies. However, absence a simple, fast efficient delivery method into primary cells been limiting progress CRISPR/Cas9-based Here, we describe an optimized protocol for iTOP-mediated various cells, including T induced pluripotent stem (hiPSCs), Jurkat, ARPE-19 HEK293 cells. We compare iTOP to other methods, such as electroporation lipofection, evaluate corresponding gene-editing efficiencies post-treatment viabilities. demonstrate that gene editing achieved by is 40–95 % depending on type, while post-iTOP viability remains high range 70–95 %. Collectively, present workflow high-throughput effective engineer difficult-to-transduce believe technology® could contribute novel