title: Genome editing of isogenic human induced pluripotent stem cells recapitulates long QT phenotype for drug testing.
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.
  
  
  OBJECTIVE
  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.
authors.affiliation: 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.
authors.forename: Yongming
authors.initials: Y
authors.lastname: Wang
authors.affiliation: 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.
authors.forename: Ping
authors.initials: P
authors.lastname: Liang
authors.affiliation: 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.
authors.forename: Feng
authors.initials: F
authors.lastname: Lan
authors.affiliation: 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.
authors.forename: Haodi
authors.initials: H
authors.lastname: Wu
authors.affiliation: Departments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, California.
authors.forename: Leszek
authors.initials: L
authors.lastname: Lisowski
authors.affiliation: 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.
authors.forename: Mingxia
authors.initials: M
authors.lastname: Gu
authors.affiliation: 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.
authors.forename: Shijun
authors.initials: S
authors.lastname: Hu
authors.affiliation: Departments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, California.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Kay
authors.affiliation: Sangamo Biosciences, Richmond, California.
authors.forename: Fyodor D
authors.initials: FD
authors.lastname: Urnov
authors.affiliation: Rambam Medical Center, Technion, Israel Institute of Technology, Haifa, Israel.
authors.forename: Rami
authors.initials: R
authors.lastname: Shinnawi
authors.affiliation: Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California.
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Gold
authors.affiliation: Rambam Medical Center, Technion, Israel Institute of Technology, Haifa, Israel.
authors.forename: Lior
authors.initials: L
authors.lastname: Gepstein
authors.affiliation: 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.
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Wu
mesh_terms: D000889:Anti-Arrhythmia Agents / Q000627:therapeutic use*; D002454:Cell Differentiation; D054338:Cell Transdifferentiation; D002478:Cells, Cultured; D016678:Genome; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000473:pathology*; D008133:Long QT Syndrome / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D018408:Patch-Clamp Techniques
chemical_list: D000889:Anti-Arrhythmia Agents
keywords: disease models; drug testing; genome editing; long QT syndrome; stem cells
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