Here, we describe a detailed method for seamless gene editing in human pluripotent stem cells using a piggyBac-based donor plasmid and the Cas9 nickase mutant. Two point mutations were introduced into exon 8 of the hepatocyte nuclear factor 4 alpha (HNF4α) locus in human embryonic stem cells (hESCs).
Wang, Y., Smith, A. J. H., Hay, D. C. Introducing Point Mutations into Human Pluripotent Stem Cells Using Seamless Genome Editing. J. Vis. Exp. (159), e61152, doi:10.3791/61152 (2020).