Targeted genome engineering using designer nucleases: State of the art and practical guidance for application in human pluripotent stem cells

Sylvia Merkert; Ulrich Martin

doi:10.1016/j.scr.2016.02.027

Targeted genome engineering using designer nucleases: State of the art and practical guidance for application in human pluripotent stem cells

Stem Cell Res. 2016 Mar;16(2):377-86. doi: 10.1016/j.scr.2016.02.027. Epub 2016 Feb 11.

Authors

Sylvia Merkert¹, Ulrich Martin²

Affiliations

¹ 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.
² 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.

PMID: 26921872
DOI: 10.1016/j.scr.2016.02.027

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.

Keywords: CRISPR/Cas9; Human pluripotent stem cells; TALEN; ZFN.

Publication types

Research Support, Non-U.S. Gov't
Review

MeSH terms

Deoxyribonucleases / genetics*
Gene Knockout Techniques
Genetic Engineering*
Homologous Recombination
Humans
Oligonucleotides / metabolism
Pluripotent Stem Cells / cytology*
Pluripotent Stem Cells / metabolism

Substances

Oligonucleotides
Deoxyribonucleases