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Development of a novel knockout model of retinitis pigmentosa using Pde6b-knockout Long–Evans rats

Although rats with melanin-pigmentated retinal pigment epithelial (RPE) cells are physiologically more appropriate models for human eye research than their albino counterparts, reliable models from the former strain are not available to study retinal degeneration. Here, we describe the development o...

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Autores principales: Yang, Jee Myung, Kim, Bora, Kwak, Jiehoon, Lee, Min Kyung, Kim, Jeong Hoon, Baek, In-Jeoung, Sung, Young Hoon, Lee, Joo Yong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9532504/
https://www.ncbi.nlm.nih.gov/pubmed/36213678
http://dx.doi.org/10.3389/fmed.2022.909182
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author Yang, Jee Myung
Kim, Bora
Kwak, Jiehoon
Lee, Min Kyung
Kim, Jeong Hoon
Baek, In-Jeoung
Sung, Young Hoon
Lee, Joo Yong
author_facet Yang, Jee Myung
Kim, Bora
Kwak, Jiehoon
Lee, Min Kyung
Kim, Jeong Hoon
Baek, In-Jeoung
Sung, Young Hoon
Lee, Joo Yong
author_sort Yang, Jee Myung
collection PubMed
description Although rats with melanin-pigmentated retinal pigment epithelial (RPE) cells are physiologically more appropriate models for human eye research than their albino counterparts, reliable models from the former strain are not available to study retinal degeneration. Here, we describe the development of a novel Pde6b-knockout Long–Evans (LE Pde6b KO) rat model that recapitulates key features of human retinitis pigmentosa (RP). After the generation of the Pde6b-knockout Sprague–Dawley rats with the CRISPR-Cpf1 system, the LE rat was back-crossed over 5 generations to develop the pigmented LE Pde6b KO strain. Interestingly, LE Pde6b KO displayed well-developed bone-spicule pigmentation; a hallmark of fundus in patients with RP which cannot be observed in non-pigmented albino rats. Moreover, the rat model showed progressive thinning of the retina, which was evident by intravital imaging with optical coherence tomography. Histologically, significant atrophy was observed in the outer nuclear layer. Functionally, LE Pde6b KO presented a marked decrease of amplitude level during electroretinogram testing, demonstrating significant loss of visual function. Therefore, these findings suggest that the LE Pde6b KO model robustly recapitulates the hallmark phenotype of RP. We believe that the LE Pde6b KO model may be used effectively for preclinical translational research to further study retinal degeneration.
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spelling pubmed-95325042022-10-06 Development of a novel knockout model of retinitis pigmentosa using Pde6b-knockout Long–Evans rats Yang, Jee Myung Kim, Bora Kwak, Jiehoon Lee, Min Kyung Kim, Jeong Hoon Baek, In-Jeoung Sung, Young Hoon Lee, Joo Yong Front Med (Lausanne) Medicine Although rats with melanin-pigmentated retinal pigment epithelial (RPE) cells are physiologically more appropriate models for human eye research than their albino counterparts, reliable models from the former strain are not available to study retinal degeneration. Here, we describe the development of a novel Pde6b-knockout Long–Evans (LE Pde6b KO) rat model that recapitulates key features of human retinitis pigmentosa (RP). After the generation of the Pde6b-knockout Sprague–Dawley rats with the CRISPR-Cpf1 system, the LE rat was back-crossed over 5 generations to develop the pigmented LE Pde6b KO strain. Interestingly, LE Pde6b KO displayed well-developed bone-spicule pigmentation; a hallmark of fundus in patients with RP which cannot be observed in non-pigmented albino rats. Moreover, the rat model showed progressive thinning of the retina, which was evident by intravital imaging with optical coherence tomography. Histologically, significant atrophy was observed in the outer nuclear layer. Functionally, LE Pde6b KO presented a marked decrease of amplitude level during electroretinogram testing, demonstrating significant loss of visual function. Therefore, these findings suggest that the LE Pde6b KO model robustly recapitulates the hallmark phenotype of RP. We believe that the LE Pde6b KO model may be used effectively for preclinical translational research to further study retinal degeneration. Frontiers Media S.A. 2022-09-21 /pmc/articles/PMC9532504/ /pubmed/36213678 http://dx.doi.org/10.3389/fmed.2022.909182 Text en Copyright © 2022 Yang, Kim, Kwak, Lee, Kim, Baek, Sung and Lee. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Medicine
Yang, Jee Myung
Kim, Bora
Kwak, Jiehoon
Lee, Min Kyung
Kim, Jeong Hoon
Baek, In-Jeoung
Sung, Young Hoon
Lee, Joo Yong
Development of a novel knockout model of retinitis pigmentosa using Pde6b-knockout Long–Evans rats
title Development of a novel knockout model of retinitis pigmentosa using Pde6b-knockout Long–Evans rats
title_full Development of a novel knockout model of retinitis pigmentosa using Pde6b-knockout Long–Evans rats
title_fullStr Development of a novel knockout model of retinitis pigmentosa using Pde6b-knockout Long–Evans rats
title_full_unstemmed Development of a novel knockout model of retinitis pigmentosa using Pde6b-knockout Long–Evans rats
title_short Development of a novel knockout model of retinitis pigmentosa using Pde6b-knockout Long–Evans rats
title_sort development of a novel knockout model of retinitis pigmentosa using pde6b-knockout long–evans rats
topic Medicine
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9532504/
https://www.ncbi.nlm.nih.gov/pubmed/36213678
http://dx.doi.org/10.3389/fmed.2022.909182
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