Correction of NR2E3 Associated Enhanced S-cone Syndrome Patient-specific iPSCs using CRISPR-Cas9

Genes (Basel). 2019 Apr 5;10(4):278. doi: 10.3390/genes10040278.


Enhanced S-cone syndrome (ESCS) is caused by recessive mutations in the photoreceptor cell transcription factor NR2E3. Loss of NR2E3 is characterized by repression of rod photoreceptor cell gene expression, over-expansion of the S-cone photoreceptor cell population, and varying degrees of M- and L-cone photoreceptor cell development. In this study, we developed a CRISPR-based homology-directed repair strategy and corrected two different disease-causing NR2E3 mutations in patient-derived induced pluripotent stem cells (iPSCs) generated from two affected individuals. In addition, one patient's iPSCs were differentiated into retinal cells and NR2E3 transcription was evaluated in CRISPR corrected and uncorrected clones. The patient's c.119-2A>C mutation caused the inclusion of a portion of intron 1, the creation of a frame shift, and generation of a premature stop codon. In summary, we used a single set of CRISPR reagents to correct different mutations in iPSCs generated from two individuals with ESCS. In doing so we demonstrate the advantage of using retinal cells derived from affected patients over artificial in vitro model systems when attempting to demonstrate pathophysiologic mechanisms of specific mutations.

Keywords: Enhanced S-Cone Syndrome (ESCS); NR2E3; clustered regularly interspaced short palindromic repeats (CRISPR); homology-directed repair (HDR); induced pluripotent stem cell (iPSC).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems / genetics
  • Cell Differentiation / genetics
  • Codon, Nonsense / genetics
  • Eye Diseases, Hereditary / genetics*
  • Eye Diseases, Hereditary / pathology
  • Eye Diseases, Hereditary / therapy*
  • Gene Expression Regulation, Developmental / genetics
  • Genetic Predisposition to Disease
  • Genetic Therapy*
  • Humans
  • Induced Pluripotent Stem Cells / metabolism
  • Induced Pluripotent Stem Cells / transplantation
  • Mutation
  • Orphan Nuclear Receptors / genetics*
  • Orphan Nuclear Receptors / therapeutic use
  • Retinal Degeneration / genetics*
  • Retinal Degeneration / pathology
  • Retinal Degeneration / therapy*
  • Retinal Rod Photoreceptor Cells / metabolism
  • Retinal Rod Photoreceptor Cells / pathology
  • Vision Disorders / genetics*
  • Vision Disorders / pathology
  • Vision Disorders / therapy*


  • Codon, Nonsense
  • NR2E3 protein, human
  • Orphan Nuclear Receptors

Supplementary concepts

  • Enhanced S-Cone Syndrome