A versatile bulk electrotransfection protocol for mouse embryonic fibroblast and iPS cells : [Preprint]

Abstract A square-wave pulsing protocol was developed using OptiMEM-GlutaMAX for high efficient transfection of mouse embryonic fibroblast and induced pluripotency stem cells. The protocol was very efficient for plasmid size ranging from 6.2 to 13.5 kb. Electroporated MFP488-labeled oligonucleotides were detected only in the cell cytoplasm, while the onset of transgene expression was during the first 4 h post-electroporation. A high rate of Venus KO cells was produced using indels as well as targeted deletion by electrotransfection of CRISPR/Cas9 plasmids. In conclusion, this plasmid electrotransfection protocol is straight-forward, cost-effective, and efficient for CRISPRing mouse primary cells.