Isolation of single-base genome-edited human iPS cells without antibiotic selection
Introduction Genome editing tools including TALENs and the CRISPR/ Cas9 system have revolutionized our ability to edit the genome of any cell, including human induced pluripotent stem cells (iPSCs). Sequence-specific nucleases induce double-strand breaks or nicks at target sites, activating the DNA repair pathways of non-homologous end joining (NHEJ) and homology-directed repair (HDR). NHEJ produces small insertions or deletions (indels) and is useful for disrupting gene function. HDR can induce precise gene repair of one to thousands of base pairs in the presence of a homologous donor molecule, allowing for correction of point mutations and introduction of exogenous sequences. Genome editing is an increasingly common part of the molecular biologist's toolkit and is being actively developed for therapeutic indications.