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
- Deoxyribonucleases/*genetics
- Gene Knockout Techniques
- *Genetic Engineering
- Homologous Recombination
- Humans
- Oligonucleotides/metabolism
- Pluripotent Stem Cells/*cytology/metabolism
- CRISPR/Cas9
- Human pluripotent stem cells
- Talen
- Zfn