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

•Gene targeting via designer nucleases in human PSCs enables various applications•Choosing the optimal methodology for a specific application is often difficult.•Useful practical recommendations for starting gene targeting in human PSCs•Overview of recent applications of designer nucleases in hPSCs

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.