Gene Therapy of Dominant CRX-Leber Congenital Amaurosis using Patient Stem Cell-Derived Retinal Organoids

Kamil Kruczek, Zepeng Qu, James Gentry, Benjamin R Fadl, Linn Gieser, Suja Hiriyanna, Zachary Batz, Mugdha Samant, Ananya Samanta, Colin J Chu, Laura Campello, Brian P Brooks, Zhijian Wu, Anand Swaroop

Research output: Contribution to journalArticle (Academic Journal)peer-review

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Mutations in the photoreceptor transcription factor gene cone-rod homeobox (CRX) lead to distinct retinopathy phenotypes, including early-onset vision impairment in dominant Leber congenital amaurosis (LCA). Using induced pluripotent stem cells (iPSCs) from a patient with CRX-I138fs48 mutation, we established an in vitro model of CRX-LCA in retinal organoids that showed defective photoreceptor maturation by histology and gene profiling, with diminished expression of visual opsins. Adeno-associated virus (AAV)-mediated CRX gene augmentation therapy partially restored photoreceptor phenotype and expression of phototransduction-related genes as determined by single-cell RNA-sequencing. Retinal organoids derived from iPSCs of a second dominant CRX-LCA patient carrying K88N mutation revealed the loss of opsin expression as a common phenotype, which was alleviated by AAV-mediated augmentation of CRX. Our studies provide a proof-of-concept for developing gene therapy of dominant CRX-LCA and other CRX retinopathies.

Original languageEnglish
Pages (from-to)252-263
Number of pages12
JournalStem Cell Reports
Issue number2
Early online date28 Jan 2021
Publication statusPublished - 9 Feb 2021


  • pluripotent stem cells
  • iPSC
  • 3-D organoids
  • retinal degeneration
  • AAV
  • therapy
  • disease modeling
  • transcription factor
  • transcriptome
  • scRNA-seq

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