June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Retinal degeneration in USH2A Mutant Rabbits Generated by CRISPR/Cas9 Technology
Author Affiliations & Notes
  • Josh Zhe
    Opthalmology, University of Michigan Michigan Medicine, Ann Arbor, Michigan, United States
  • Van Phuc Nguyen
    Opthalmology, University of Michigan Michigan Medicine, Ann Arbor, Michigan, United States
  • Jun Song
    Center for Advanced Models for Translational Sciences and Therapeutics, Ann Arbor, Michigan, United States
  • Yanxiu Li
    Center for Advanced Models for Translational Sciences and Therapeutics, Ann Arbor, Michigan, United States
  • Yifei Sun
    Center for Advanced Models for Translational Sciences and Therapeutics, Ann Arbor, Michigan, United States
  • Jie Xu
    Center for Advanced Models for Translational Sciences and Therapeutics, Ann Arbor, Michigan, United States
  • Jifeng Zhang
    Center for Advanced Models for Translational Sciences and Therapeutics, Ann Arbor, Michigan, United States
  • Y. Eugene Chen
    Center for Advanced Models for Translational Sciences and Therapeutics, Ann Arbor, Michigan, United States
  • Dongshan Yang
    Center for Advanced Models for Translational Sciences and Therapeutics, Ann Arbor, Michigan, United States
  • Yannis Mantas Paulus
    Opthalmology, University of Michigan Michigan Medicine, Ann Arbor, Michigan, United States
  • Footnotes
    Commercial Relationships   Josh Zhe None; Van Phuc Nguyen None; Jun Song None; Yanxiu Li None; Yifei Sun None; Jie Xu None; Jifeng Zhang None; Y. Eugene Chen None; Dongshan Yang None; Yannis Paulus None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 1782 – F0331. doi:
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    • Get Citation

      Josh Zhe, Van Phuc Nguyen, Jun Song, Yanxiu Li, Yifei Sun, Jie Xu, Jifeng Zhang, Y. Eugene Chen, Dongshan Yang, Yannis Mantas Paulus; Retinal degeneration in USH2A Mutant Rabbits Generated by CRISPR/Cas9 Technology. Invest. Ophthalmol. Vis. Sci. 2022;63(7):1782 – F0331.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Usher Syndrome type IIA (USH2A) is a genetic disorder characterized by progressive photoreceptor degeneration due to retinitis pigmentosa (RP) and sensorineural deafness. Currently, there exists no therapy to stop or reverse the progressive photoreceptor death in USH2A. In order to understand and treat retinal degeneration in patients, it is essential to create a USH2A animal disease model. Since the Usherin protein is highly conserved in humans and rabbits, rabbits make a great model for USH2A. Furthermore, rabbit eyes have similar anatomical features to human eyes. In this study, we successfully created a USH2A knockout rabbit using CRISPR/Cas9 technology.

Methods : Rabbit embryos were injected with a mixture of 150ng/μL Cas9 mRNA and 50 ng/μL sgRNA at a dose of 2-5 pL. The guide RNA targets the rabbit USH2A gene on exon 12. Each recipient female rabbit received twenty to thirty injected embryos through embryo transfer surgery. Derived founder rabbits were bred to be homozygous for the USH2A knockout. The progression of retinal degeneration is then evaluated for up to 22 months using fundus photography, fundus autofluorescence (FAF), fluorescein angiography (FA), OCT, electroretinography (ERG), and slit lamp examination.

Results : In the three biallelic mutant rabbits, FAF spots were detected in the mid-periphery of the retina, and OCT images showed hyper-reflective disruptions of the photoreceptor layer. ERG signals of both rod and cone function were reduced in the USH2A mutant rabbits at 7 months and worsened at 15-22 months, indicating progressive retinal photoreceptor degeneration.

Conclusions : Due to similar anatomical features and disease progression as noted in humans, our USH2A rabbit model is a useful tool for studying retinal degeneration in Usher syndrome.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

 

In vivo imaging of retinal degeneration in USH2A rabbit model: (a) Color fundus photographs, fundus autofluorescence (FAF), fluorescein (FA) images, and indocyanine green angiography (ICGA) achieved from control (top) and the USH2A mutant rabbit at 5-month-old (bottom). (b) B-scan OCT images at 5 months old. (c) Electroretinogram (ERG) rod response of control (black) and USH2A (white) groups.

In vivo imaging of retinal degeneration in USH2A rabbit model: (a) Color fundus photographs, fundus autofluorescence (FAF), fluorescein (FA) images, and indocyanine green angiography (ICGA) achieved from control (top) and the USH2A mutant rabbit at 5-month-old (bottom). (b) B-scan OCT images at 5 months old. (c) Electroretinogram (ERG) rod response of control (black) and USH2A (white) groups.

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