Abstract LB-043: Generation of clonal CRISPR/Cas9-edited human iPSC derived cellular models and its applications in physiologically relevant assays

Induced pluripotent stem cell (iPSC)-derived cells are widely used in disease models, safety testing and phenotypic and functional assays. The CRISPR/Cas9 system has revolutionized genome editing thus enabling rapid generation of disease models. Cutting edge technologies like genome editing paired with iPSC-derived cell models have reshaped our approach to disease modeling in vitro. Although, CRISPR/Cas9 has had huge impact on generation of complex cellular models, there are challenges in replicating disease-in-a-dish. One of the challenges is the implementation of genome editing tools in iPSC, particularly issues associated with editing tools delivery, cell recovery and clonal isolation of genome-edited cells. Here, we describe a workflow that facilitates the generation and isolation of clonal CRISPR-edited iPSCs through the use of a novel toolset. We observed high genome editing efficiency using an iPSC line that stably expresses the Cas9 protein and obtained improved survival of the iPSCs after delivery of the editing tools through culture in StemFlex™ medium, a novel PSC culture system. Furthermore, we have demonstrated that single edited iPSCs can be isolated through the combination of an optimized cell sorting method and expanded as single cell clones with reagents that support single cell survival. Through this workflow we have generated several iPSC lines carrying disease relevant SNP mutations. These tools and workflows not only contribute to improved success in the derivation of homogenous genome edited human iPSC clones but also provide novel alternatives to study disease-causing mutations in vitro. Additionally, we have also tested these cell models in commonly used cell health assays. These cell models can be further used to generate organoids to mimic conditions found in tissues. iPSC- derived cell models not only permit the study of disease mechanism in the cell types specifically affected in the disease but also deepen our understanding of the molecular mechanisms and in turn help development of novel therapeutics. Citation Format: Chetana M. Revankar, Erik Willems, Rex Lacambacal, Tufan gokirmak, Jordan Dizon, Jacquelyn Webb, Raquel Vega, David Piper. Generation of clonal CRISPR/Cas9-edited human iPSC derived cellular models and its applications in physiologically relevant assays [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-043.