Developing a robust pipeline for high throughput CRISPR/Cas-mediated gene editing in human induced pluripotent stem cells
Human induced pluripotent stem cell (hiPSC) technology has propelled the stem cell biology field; the ability of iPSCs to give rise to almost any cell type in the body offers opportunity for modelling diseases, drug discovery and cell therapy development. The capacity to genetically define iPSC-derived human disease models using CRISPR/Cas endonucleases significantly enhances their potential application. We have previously developed an automated pipeline to generate other genetically modified mammalian cell lines in a high throughput manner. We aimed to adapt this pipeline for CRISPR/Cas mediated gene editing of iPSCs to meet an increasing demand for custom iPSC manufacture and generation of complex disease models. Gene editing pluripotent stem cells can be challenging due to their sensitivity to spontaneous differentiation and their reported low editing efficiency. Therefore, we set out to develop an improved iPSC gene editing protocol, which would deliver an appropriate balance between efficiency and maintained cellular homeostasis.
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