Human induced pluripotent stem cells (hiPSCs) offer an unprecedented opportunity for in vitro disease modeling and for personalized cell replacement therapy1. Applications of iPSCs have been greatly expanded by the advent of genome editing, in which the genomic sequence at a target site is altered by insertion or deletion (“indel”) mutations, or by introduction of precisely programmed (“knockin”) modifications2. Here we present a highly efficient and reproducible protocol to edit the genome of hiPSCs through the combined use of the CRISPR/Cas9 RNA-guided nuclease and piggyBac transposase3–5. This protocol is best suited to applications in which a common starting cell line is edited many different times to yield isogenic daughter cell lines that differ by the introduced mutations.
