title: Targeted genome engineering using designer nucleases: State of the art and practical guidance for application in human pluripotent stem cells.
abstract: Within the last years numerous publications successfully applied sequence specific designer nucleases for genome editing in human PSCs. However, despite this abundance of reports together with the rapid development and improvement accomplished with the technology, it is still difficult to choose the optimal methodology for a specific application of interest. With focus on the most suitable approach for specific applications, we present a practical guidance for successful gene editing in human PSCs using designer nucleases. We discuss experimental considerations, limitations and critical aspects which will guide the investigator for successful implementation of this technology.
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany; REBIRTH - Cluster of Excellence, Hannover Medical School, Germany; Biomedical Research in Endstage and Obstructive Lung Disease (BREATH), Member of the German Center for Lung Research (DZL), Germany.
authors.forename: Sylvia
authors.initials: S
authors.lastname: Merkert
authors.affiliation: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, 30625 Hannover, Germany; REBIRTH - Cluster of Excellence, Hannover Medical School, Germany; Biomedical Research in Endstage and Obstructive Lung Disease (BREATH), Member of the German Center for Lung Research (DZL), Germany. Electronic address: martin.ulrich@mh-hannover.de.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Martin
mesh_terms: D003851:Deoxyribonucleases / Q000235:genetics*; D055786:Gene Knockout Techniques; D005818:Genetic Engineering; D059765:Homologous Recombination; D006801:Humans; D009841:Oligonucleotides / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D009841:Oligonucleotides; D003851:Deoxyribonucleases
keywords: CRISPR/Cas9; Human pluripotent stem cells; TALEN; ZFN