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J Am Coll Cardiol. 2014 Aug 5;64(5):451-9. doi: 10.1016/j.jacc.2014.04.057.

Genome editing of isogenic human induced pluripotent stem cells recapitulates long QT phenotype for drug testing.

Author information

1
Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, China.
2
Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California.
3
Departments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, California.
4
Sangamo Biosciences, Richmond, California.
5
Rambam Medical Center, Technion, Israel Institute of Technology, Haifa, Israel.
6
Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California.
7
Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California. Electronic address: joewu@stanford.edu.

Abstract

BACKGROUND:

Human induced pluripotent stem cells (iPSCs) play an important role in disease modeling and drug testing. However, the current methods are time-consuming and lack an isogenic control.

OBJECTIVES:

This study sought to establish an efficient technology to generate human PSC-based disease models with isogenic control.

METHODS:

The ion channel genes KCNQ1 and KCNH2 with dominant negative mutations causing long QT syndrome types 1 and 2, respectively, were stably integrated into a safe harbor AAVS1 locus using zinc finger nuclease technology.

RESULTS:

Patch-clamp recording revealed that the edited iPSC-derived cardiomyocytes (iPSC-CMs) displayed characteristic long QT syndrome phenotype and significant prolongation of the action potential duration compared with the unedited control cells. Finally, addition of nifedipine (L-type calcium channel blocker) or pinacidil (KATP-channel opener) shortened the action potential duration of iPSC-CMs, confirming the validity of isogenic iPSC lines for drug testing in the future.

CONCLUSIONS:

Our study demonstrates that iPSC-CM-based disease models can be rapidly generated by overexpression of dominant negative gene mutants.

KEYWORDS:

disease models; drug testing; genome editing; long QT syndrome; stem cells

PMID:
25082577
PMCID:
PMC4149735
DOI:
10.1016/j.jacc.2014.04.057
[Indexed for MEDLINE]
Free PMC Article

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