DATE: June 19, 2019
TIME: 11:00am PDT, 2:00pm EDT
Human induced pluripotent stem cells (hiPSCs) have been globally recognized as a multipurpose research tool for modeling human disease and biology, screening and developing potential therapeutic drugs, and implementing cell and gene therapies. The ability to differentiate human iPSCs into any cell type supports the study of biology and disease in these specified cells in vitro. The emergence of genome editing tools, inlcuding the CRISPR/Cas9 system or TALENs, enable genetic modification of these cells; such as introduction of single base changes or inserting reporters or bio-sensors, which can be used to study the effects of genetic differences or biological functions in the desired cell type. Given the challenges with genome editing efficiency, cell survival and clonal isolation, we have developed a number of reagents and processes to dramatically improve the success rate and timelines for a genome editing experiment in hiPSCs. Key areas of the genome editing workflow that have been addressed include the genome editing tools themselves, the delivery methods and the maintenance of healthy hiPSC cultures during these stressful manipulations. Using the generation of disease models relevant to cardiac and neuronal disease to explore and identify the best workflow for the genome editing process in hiPSC, we built a reliable approach that reproducibly supports the generation of hiPSC lines carrying small mutations such as SNPs or small deletions. Subsequent studies of the disease relevant cell types then identified cellular phenotypes that corroborated with those previously identified in patient-derived hiPSC-based models. In summary, we detail advances with tools, reagents and protocols that facilitate the genome editing workflow in hiPSC and demonstrate that the use of such tools can be readily implemented to generate hiPSC-derived disease models and reporter lines.
- Selecting the right tools are key for successful genome editing in PSC
- Successful clonal isolation of PSC heavily depends on medium choice.
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