Use of CRISPR/Cas ribonucleoproteins for high throughput gene editing of induced pluripotent stem cells.
Qi Wang, Sueanne Chear, Kristof Wing, David Stellon, Minh Thuan Nguyen Tran, Jana Talbot, Alice Pébay, Alex W Hewitt, Anthony L Cook
Methods | Published : 2021
Induced pluripotent stem cells (iPSCs) have become widely used for disease modelling, particularly with regard to predisposing genetic risk factors and causal gene variants. Alongside this, technologies such as the CRISPR/Cas system have been adapted to enable programmable gene editing in human cells. When combined, CRISPR/Cas gene editing of donor-specific iPSC to generate isogenic cell lines that differ only at specific gene variants provides a powerful model with which to investigate genetic variants associated with diseases affecting many organs, including the brain and eye. Here we describe our optimized protocol for using CRISPR/Cas ribonucleoproteins to edit disease causing gene varia..View full abstract