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Updated: Oct 27, 2025

Establishment of Genome-edited Human Pluripotent Stem Cell Lines: From Targeting to Isolation
Published on: February 2, 2016
Genome Editing in Human Induced Pluripotent Stem Cells (hiPSCs)
Shuichiro Higo1,2, Shungo Hikoso3, Shigeru Miyagawa4
1Department of Medical Therapeutics for Heart Failure, Osaka University Graduate School of Medicine, Osaka, Japan. higo-s@cardiology.med.osaka-u.ac.jp.
Abstract:
Cardiomyocytes differentiated from human induced pluripotent stem cells (hiPSCs) are powerful tools for elucidating the pathology behind inherited cardiomyopathies. Genome editing technologies enable targeted genome replacement and the generation of isogenic hiPSCs, allowing investigators to precisely determine the roles of identified mutations. Here, we describe a protocol to obtain isogenic hiPSCs with the corrected allele via homology-directed repair (HDR) using CRISPR/Cas9 genome editing under feeder-free conditions. Seeding hiPSCs in a 24-well plate and conducting the initial evaluation using direct genomic sequencing after 1 week is cost- and time-effective. Following optimization of the protocol, sequence confirmation of the corrected HDR clone is completed within 21 days.
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