Enhanced genome editing in human iPSCs with CRISPR-CAS9 by co-targeting ATP1a1.

ABSTRACT: Genome editing in human induced pluripotent stem cells (iPSCs) provides the potential for disease modeling and cell therapy. By generating iPSCs with specific mutations, researchers can differentiate the modified cells to their lineage of interest for further investigation. However, the low efficiency of targeting in iPSCs has hampered the application of genome editing. In this study we used a CRISPR-Cas9 system that introduces a specific point substitution into the sequence of the Na⁺/K⁺-ATPase subunit ATP1A1. The introduced mutation confers resistance to cardiac glycosides, which can then be used to select successfully targeted cells. Using this system, we introduced different formats of donor DNA for homology-directed repair (HDR), including single-strand DNAs, dou

SUBMITTER: Liu JT 

PROVIDER: S-EPMC7197401 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature