title: Genome Editing in Human Pluripotent Stem Cells.
abstract: Genome editing in human pluripotent stem cells (hPSCs) enables the generation of reporter lines and knockout cell lines. Zinc finger nucleases, transcription activator-like effector nucleases (TALENs), and CRISPR/Cas9 technology have recently increased the efficiency of proper gene editing by creating double strand breaks (DSB) at defined sequences in the human genome. These systems typically use plasmids to transiently transcribe nucleases within the cell. Here, we describe the process for preparing hPSCs for transient expression of nucleases via electroporation and subsequent analysis to create genetically modified stem cell lines.
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, 1550 Engineering Drive, Madison, WI, 53706, USA.|Wisconsin Institute for Discovery, University of Wisconsin-Madison, 330 N. Orchard Street, Madison, WI, 53715, USA.
authors.forename: Jared
authors.initials: J
authors.lastname: Carlson-Stevermer
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, 1550 Engineering Drive, Madison, WI, 53706, USA. ksaha@wisc.edu.|Wisconsin Institute for Discovery, University of Wisconsin-Madison, 330 N. Orchard Street, Madison, WI, 53715, USA. ksaha@wisc.edu.
authors.forename: Krishanu
authors.initials: K
authors.lastname: Saha
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics; D002460:Cell Line; D018274:Electroporation / Q000379:methods; D000072669:Gene Editing / Q000379:methods; D005818:Genetic Engineering / Q000379:methods; D015894:Genome, Human / Q000235:genetics; D006801:Humans; D039904:Pluripotent Stem Cells / Q000502:physiology*; D000069896:Transcription Activator-Like Effector Nucleases / Q000235:genetics; D000075804:Zinc Finger Nucleases / Q000235:genetics
chemical_list: D000069896:Transcription Activator-Like Effector Nucleases; D000075804:Zinc Finger Nucleases
keywords: CRISPR/Cas9; Electroporation; Genome editing; Genomics