Cargando…
In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway
Although Cas9-mediated genome editing has been widely used to engineer alleles in animal models of human inherited diseases, very few homology-directed repair (HDR)–based genetic editing systems have been established in postnatal mouse models for effective and lasting phenotypic rescue. Here, we dev...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Association for the Advancement of Science
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6469935/ https://www.ncbi.nlm.nih.gov/pubmed/31001583 http://dx.doi.org/10.1126/sciadv.aav3335 |
_version_ | 1783411706319339520 |
---|---|
author | Cai, Yuan Cheng, Tianlin Yao, Yichuan Li, Xiao Ma, Yuqian Li, Lingyun Zhao, Huan Bao, Jin Zhang, Mei Qiu, Zilong Xue, Tian |
author_facet | Cai, Yuan Cheng, Tianlin Yao, Yichuan Li, Xiao Ma, Yuqian Li, Lingyun Zhao, Huan Bao, Jin Zhang, Mei Qiu, Zilong Xue, Tian |
author_sort | Cai, Yuan |
collection | PubMed |
description | Although Cas9-mediated genome editing has been widely used to engineer alleles in animal models of human inherited diseases, very few homology-directed repair (HDR)–based genetic editing systems have been established in postnatal mouse models for effective and lasting phenotypic rescue. Here, we developed an HDR-based Cas9/RecA system to precisely correct Pde6b mutation with increased HDR efficiency in postnatal rodless (rd1) mice, a retinitis pigmentosa (RP) mutant model characterized by photoreceptor degeneration and loss of vision. The Cas9/RecA system incorporated Cas9 endonuclease enzyme to generate double-strand breaks (DSBs) and bacterial recombinase A (RecA) to increase homologous recombination. Our data revealed that Cas9/RecA treatment significantly promoted the survival of both rod and cone photoreceptors, restored the expression of PDE6B in rod photoreceptors, and enhanced the visual functions of rd1 mice. Thus, this study provides a precise therapeutic strategy for RP and other genetic diseases. |
format | Online Article Text |
id | pubmed-6469935 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | American Association for the Advancement of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-64699352019-04-18 In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway Cai, Yuan Cheng, Tianlin Yao, Yichuan Li, Xiao Ma, Yuqian Li, Lingyun Zhao, Huan Bao, Jin Zhang, Mei Qiu, Zilong Xue, Tian Sci Adv Research Articles Although Cas9-mediated genome editing has been widely used to engineer alleles in animal models of human inherited diseases, very few homology-directed repair (HDR)–based genetic editing systems have been established in postnatal mouse models for effective and lasting phenotypic rescue. Here, we developed an HDR-based Cas9/RecA system to precisely correct Pde6b mutation with increased HDR efficiency in postnatal rodless (rd1) mice, a retinitis pigmentosa (RP) mutant model characterized by photoreceptor degeneration and loss of vision. The Cas9/RecA system incorporated Cas9 endonuclease enzyme to generate double-strand breaks (DSBs) and bacterial recombinase A (RecA) to increase homologous recombination. Our data revealed that Cas9/RecA treatment significantly promoted the survival of both rod and cone photoreceptors, restored the expression of PDE6B in rod photoreceptors, and enhanced the visual functions of rd1 mice. Thus, this study provides a precise therapeutic strategy for RP and other genetic diseases. American Association for the Advancement of Science 2019-04-17 /pmc/articles/PMC6469935/ /pubmed/31001583 http://dx.doi.org/10.1126/sciadv.aav3335 Text en Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC). http://creativecommons.org/licenses/by-nc/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license (http://creativecommons.org/licenses/by-nc/4.0/) , which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited. |
spellingShingle | Research Articles Cai, Yuan Cheng, Tianlin Yao, Yichuan Li, Xiao Ma, Yuqian Li, Lingyun Zhao, Huan Bao, Jin Zhang, Mei Qiu, Zilong Xue, Tian In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway |
title | In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway |
title_full | In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway |
title_fullStr | In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway |
title_full_unstemmed | In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway |
title_short | In vivo genome editing rescues photoreceptor degeneration via a Cas9/RecA-mediated homology-directed repair pathway |
title_sort | in vivo genome editing rescues photoreceptor degeneration via a cas9/reca-mediated homology-directed repair pathway |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6469935/ https://www.ncbi.nlm.nih.gov/pubmed/31001583 http://dx.doi.org/10.1126/sciadv.aav3335 |
work_keys_str_mv | AT caiyuan invivogenomeeditingrescuesphotoreceptordegenerationviaacas9recamediatedhomologydirectedrepairpathway AT chengtianlin invivogenomeeditingrescuesphotoreceptordegenerationviaacas9recamediatedhomologydirectedrepairpathway AT yaoyichuan invivogenomeeditingrescuesphotoreceptordegenerationviaacas9recamediatedhomologydirectedrepairpathway AT lixiao invivogenomeeditingrescuesphotoreceptordegenerationviaacas9recamediatedhomologydirectedrepairpathway AT mayuqian invivogenomeeditingrescuesphotoreceptordegenerationviaacas9recamediatedhomologydirectedrepairpathway AT lilingyun invivogenomeeditingrescuesphotoreceptordegenerationviaacas9recamediatedhomologydirectedrepairpathway AT zhaohuan invivogenomeeditingrescuesphotoreceptordegenerationviaacas9recamediatedhomologydirectedrepairpathway AT baojin invivogenomeeditingrescuesphotoreceptordegenerationviaacas9recamediatedhomologydirectedrepairpathway AT zhangmei invivogenomeeditingrescuesphotoreceptordegenerationviaacas9recamediatedhomologydirectedrepairpathway AT qiuzilong invivogenomeeditingrescuesphotoreceptordegenerationviaacas9recamediatedhomologydirectedrepairpathway AT xuetian invivogenomeeditingrescuesphotoreceptordegenerationviaacas9recamediatedhomologydirectedrepairpathway |