Reversal of Phenotypic Abnormalities by CRISPR/Cas9-Mediated Gene Correction in Huntington Disease Patient-Derived Induced Pluripotent Stem Cells

Stem Cell Reports. 2017 Mar 14;8(3):619-633. doi: 10.1016/j.stemcr.2017.01.022. Epub 2017 Feb 23.


Huntington disease (HD) is a dominant neurodegenerative disorder caused by a CAG repeat expansion in HTT. Here we report correction of HD human induced pluripotent stem cells (hiPSCs) using a CRISPR-Cas9 and piggyBac transposon-based approach. We show that both HD and corrected isogenic hiPSCs can be differentiated into excitable, synaptically active forebrain neurons. We further demonstrate that phenotypic abnormalities in HD hiPSC-derived neural cells, including impaired neural rosette formation, increased susceptibility to growth factor withdrawal, and deficits in mitochondrial respiration, are rescued in isogenic controls. Importantly, using genome-wide expression analysis, we show that a number of apparent gene expression differences detected between HD and non-related healthy control lines are absent between HD and corrected lines, suggesting that these differences are likely related to genetic background rather than HD-specific effects. Our study demonstrates correction of HD hiPSCs and associated phenotypic abnormalities, and the importance of isogenic controls for disease modeling using hiPSCs.

Keywords: CHCHD2; Huntington disease; disease modeling; disease phenotypes; genetic editing; human induced pluripotent stem cell (hiPSC); mitochondrial dysfunction; neurodegenerative disorders; transcriptional dysrgulation.

MeSH terms

  • CRISPR-Cas Systems*
  • Cell Differentiation / genetics
  • Cell Line
  • Cell Self Renewal / genetics
  • DNA-Binding Proteins
  • Electrophysiological Phenomena / genetics
  • Gene Editing*
  • Gene Expression Regulation, Developmental
  • Gene Targeting
  • Humans
  • Huntington Disease / genetics*
  • Huntington Disease / metabolism*
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism*
  • Mitochondria / genetics
  • Mitochondria / metabolism
  • Mitochondrial Proteins / genetics
  • Neurons / cytology
  • Neurons / metabolism
  • Phenotype*
  • Transcription Factors / genetics


  • CHCHD2 protein, human
  • DNA-Binding Proteins
  • Mitochondrial Proteins
  • Transcription Factors