title: Genome Editing in Human Pluripotent Stem Cells: Approaches, Pitfalls, and Solutions.
abstract: Human pluripotent stem cells (hPSCs) with knockout or mutant alleles can be generated using custom-engineered nucleases. Transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 nucleases are the most commonly employed technologies for editing hPSC genomes. In this Protocol Review, we provide a brief overview of custom-engineered nucleases in the context of gene editing in hPSCs with a focus on the application of TALENs and CRISPR/Cas9. We will highlight the advantages and disadvantages of each method and discuss theoretical and technical considerations for experimental design.
authors.affiliation: The Collaborative Center for X-Linked Dystonia Parkinsonism, Department of Neurology, Massachusetts General Hospital, Charlestown, MA 02129, USA; Harvard Brain Science Initiative, Harvard Medical School, Boston, MA 02114, USA.
authors.forename: William T
authors.initials: WT
authors.lastname: Hendriks
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA.
authors.forename: Curtis R
authors.initials: CR
authors.lastname: Warren
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA 02114, USA. Electronic address: chad_cowan@harvard.edu.
authors.forename: Chad A
authors.initials: CA
authors.lastname: Cowan
mesh_terms: D000483:Alleles; D064113:CRISPR-Cas Systems; D004247:DNA / Q000032:analysis; D003851:Deoxyribonucleases / Q000235:genetics / Q000378:metabolism; D005821:Genetic Techniques; D005822:Genetic Vectors; D015894:Genome, Human; D005838:Genotype; D006801:Humans; D009154:Mutation; D039904:Pluripotent Stem Cells / Q000166:cytology*; D017354:Point Mutation; D012689:Sequence Homology, Nucleic Acid
chemical_list: D004247:DNA; D003851:Deoxyribonucleases