CRISPR correction of the PRKAG2 gene mutation in the patient's induced pluripotent stem cell-derived cardiomyocytes eliminates electrophysiological and structural abnormalities

Research output: Contribution to journalResearch articleContributedpeer-review


  • Ronen Ben Jehuda - , Technion-Israel Institute of Technology (Author)
  • Binyamin Eisen - , Technion-Israel Institute of Technology (Author)
  • Yuval Shemer - , Technion-Israel Institute of Technology (Author)
  • Lucy N. Mekies - , Technion-Israel Institute of Technology (Author)
  • Agnes Szantai - , Technion-Israel Institute of Technology, University of Szeged (Author)
  • Irina Reiter - , Technion-Israel Institute of Technology (Author)
  • Huanhuan Cui - , Charité – Universitätsmedizin Berlin (Author)
  • Kaomei Guan - , Deutsches Zentrum für Herz-Kreislaufforschung (DZHK), University Medical Center Göttingen, University of Göttingen (Author)
  • Shiraz Haron-Khun - , Tel Aviv University (Author)
  • Dov Freimark - , Tel Aviv University (Author)
  • Silke R. Sperling - , Charité – Universitätsmedizin Berlin, Deutsches Zentrum für Herz-Kreislaufforschung (DZHK) (Author)
  • Mihaela Gherghiceanu - , Victor Babes National Institute (Author)
  • Michael Arad - , Tel Aviv University (Author)
  • Ofer Binah - , Technion-Israel Institute of Technology (Author)


Background Mutations in the PRKAG2 gene encoding the γ-subunit of adenosine monophosphate kinase (AMPK) cause hypertrophic cardiomyopathy (HCM) and familial Wolff-Parkinson-White (WPW) syndrome. Patients carrying the R302Q mutation in PRKAG2 present with sinus bradycardia, escape rhythms, ventricular preexcitation, supraventricular tachycardia, and atrioventricular block. This mutation affects AMPK activity and increases glycogen storage in cardiomyocytes. The link between glycogen storage, WPW syndrome, HCM, and arrhythmias remains unknown. Objective The purpose of this study was to investigate the pathological changes caused by the PRKAG2 mutation. We tested the hypothesis that patient's induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) display clinical aspects of the disease. Methods Using clustered regularly interspaced short palindromic repeats (CRISPR) technology, we corrected the mutation and then generated isogenic iPSC-CMs. Action potentials were recorded from spontaneously firing and paced cardiomyocytes using the patch clamp technique. Using a microelectrode array setup, we recorded electrograms from iPSC-CMs clusters. Transmission electron microscopy was used to detect ultrastructural abnormalities in the mutated iPSC-CMs. Results PRKAG2-mutated iPSC-CMs exhibited abnormal firing patterns, delayed afterdepolarizations, triggered arrhythmias, and augmented beat rate variability. Importantly, CRISPR correction eliminated the electrophysiological abnormalities, the augmented glycogen, storage, and cardiomyocyte hypertrophy. Conclusion PRKAG2-mutated iPSC-CMs displayed functional and structural abnormalities, which were abolished by correcting the mutation in the patient's iPSCs using CRISPR technology.


Original languageEnglish
Pages (from-to)267-276
Number of pages10
JournalHeart Rhythm
Issue number2
Publication statusPublished - Feb 2018
Externally publishedYes

External IDs

Scopus 85041337297
PubMed 28917552



  • arrhythmia, clustered regularly interspaced short palindromic repeats, electrophysiology, induced pluripotent stem cell-derived cardiomyocyte, PRKAG2, Wolff-Parkinson-White syndrome