2 × 10e5
HEK293 cells were harvested, washed once in PBS, and resuspended in 20 µL
of SF nucleofection buffer (Lonza, Basel, Switzerland). 10 µL of RNP mixture (nuclease and sgRNA complex), 100 pmol of donor DNA, and cell suspension were combined in a Lonza 4D 16well strip
nucleocuvette. Reaction mixtures were electroporated using setting DS150,
incubated in the nucleocuvette at RT for 10 min, and transferred to culture
dishes containing pre-warmed media.