This protocol requires establishing a parental cell line with the stably integrated transgene, which represents an extra step compared to methods based on transient transfection coupled with cell sorting-based enrichment of transfected cells. However, the benefit of our strategy is increased consistency and higher efficiency for recovering genome edited clones once the parental line is established. Therefore we use this strategy in situations where it is desirable to make many different genome-edited cell lines from a common parent, such as when generating a series of cell lines that are isogenic except for introduced mutations. On the other hand, this strategy would be cumbersome for editing many different cell lines, such as for correction of mutations in patient-derived iPSC lines.
