Mouse Models of Achromatopsia in Addressing Temporal “Point of No Return” in Gene-Therapy
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Published:2021-07-28
Issue:15
Volume:22
Page:8069
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ISSN:1422-0067
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Container-title:International Journal of Molecular Sciences
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language:en
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Short-container-title:IJMS
Author:
Wang Nan-KaiORCID,
Liu Pei-KangORCID,
Kong Yang,
Levi Sarah R.,
Huang Wan-Chun,
Hsu Chun-Wei,
Wang Hung-Hsi,
Chen Nelson,
Tseng Yun-Ju,
Quinn Peter M. J.ORCID,
Tai Ming-Hong,
Lin Chyuan-Sheng,
Tsang Stephen H.
Abstract
Achromatopsia is characterized by amblyopia, photophobia, nystagmus, and color blindness. Previous animal models of achromatopsia have shown promising results using gene augmentation to restore cone function. However, the optimal therapeutic window to elicit recovery remains unknown. Here, we attempted two rounds of gene augmentation to generate recoverable mouse models of achromatopsia including a Cnga3 model with a knock-in stop cassette in intron 5 using Easi-CRISPR (Efficient additions with ssDNA inserts-CRISPR) and targeted embryonic stem (ES) cells. This model demonstrated that only 20% of CNGA3 levels in homozygotes derived from target ES cells remained, as compared to normal CNGA3 levels. Despite the low percentage of remaining protein, the knock-in mouse model continued to generate normal cone phototransduction. Our results showed that a small amount of normal CNGA3 protein is sufficient to form “functional” CNG channels and achieve physiological demand for proper cone phototransduction. Thus, it can be concluded that mutating the Cnga3 locus to disrupt the functional tetrameric CNG channels may ultimately require more potent STOP cassettes to generate a reversible achromatopsia mouse model. Our data also possess implications for future CNGA3-associated achromatopsia clinical trials, whereby restoration of only 20% functional CNGA3 protein may be sufficient to form functional CNG channels and thus rescue cone response.
Funder
National Institutes of Health
Schneeweiss Stem Cell Fund, New York State
Ministry of Science and Technology, Taiwan
Kaohsiung Medical University Chung-Ho Memorial Hospital
Subject
Inorganic Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Computer Science Applications,Spectroscopy,Molecular Biology,General Medicine,Catalysis
Cited by
2 articles.
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