Caspase-1 knockout disrupts pyroptosis and protects photoreceptor cells from photochemical damage
Aim: Retinal photochemical damage (RPD) plays a significant role in the development of various ocular diseases, with Caspase-1 being a key contributor. This study investigates the protective effects of Caspase-1 gene-mediated pyroptosis against RPD. Methods: Differentially expressed genes (DEGs) ass...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2024-12-01
|
| Series: | Molecular and Cellular Probes |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0890850824000434 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850249679191670784 |
|---|---|
| author | Xiaoping Yu Jiayuan Peng Qian Zhong Ailin Wu Xiaoming Deng Yanfeng Zhu |
| author_facet | Xiaoping Yu Jiayuan Peng Qian Zhong Ailin Wu Xiaoming Deng Yanfeng Zhu |
| author_sort | Xiaoping Yu |
| collection | DOAJ |
| description | Aim: Retinal photochemical damage (RPD) plays a significant role in the development of various ocular diseases, with Caspase-1 being a key contributor. This study investigates the protective effects of Caspase-1 gene-mediated pyroptosis against RPD. Methods: Differentially expressed genes (DEGs) associated with RPD were identified through the analysis of two expression profiles from the GEO database. Correlation analysis was used to pinpoint pyroptosis-related genes (PRGs) linked to RPD. A Caspase-1 knockout 661 W cell line was generated via CRISPR-Cas9 gene editing, and single-cell colonies were screened and purified. Validation of knockout cells was performed through RT-qPCR, gene sequencing, and Western blot analysis. Comparative assays on cell proliferation, intracellular reactive oxygen species (ROS), and cytotoxicity were conducted between wild-type and Caspase-1 knockout cells under light exposure. Further RT-qPCR and Western blot experiments examined changes in the mRNA and protein levels of key pyroptosis pathway components. Results: Significant alterations in Caspase-1 expression were observed among PRGs. Homozygous Caspase-1 knockout cell lines were confirmed through RT-qPCR, genomic PCR product sequencing, and Western blot analysis. Compared to wild-type 661 W cells, Caspase-1 knockout cells exhibited higher viability and proliferation rates after 24 h of light exposure, alongside reduced LDH release. The expression of downstream pyroptosis factors at both the mRNA and protein levels was markedly decreased in the knockout group. Conclusion: CRISPR/Cas9-mediated Caspase-1 knockout enhanced the resistance of 661 W cells to photochemical damage, suggesting that Caspase-1 may serve as a potential therapeutic target for RPD-related diseases. |
| format | Article |
| id | doaj-art-c1a5c601d0e3448d8e05652deec178ce |
| institution | OA Journals |
| issn | 0890-8508 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular and Cellular Probes |
| spelling | doaj-art-c1a5c601d0e3448d8e05652deec178ce2025-08-20T01:58:27ZengElsevierMolecular and Cellular Probes0890-85082024-12-017810199110.1016/j.mcp.2024.101991Caspase-1 knockout disrupts pyroptosis and protects photoreceptor cells from photochemical damageXiaoping Yu0Jiayuan Peng1Qian Zhong2Ailin Wu3Xiaoming Deng4Yanfeng Zhu5School of Medicine and Nursing, Chengdu University, 610106, Sichuan Province, China; Department of Public Health, Chengdu Medical College, 610500, Sichuan Province, China; Corresponding author. School of Medicine and Nursing, Chengdu University, 610106, Sichuan Province, China.Department of Pathology, People's Hospital of Leshan, 614000, Sichuan Province, China; Department of Basic Medical Science, Chengdu Medical College, 610500, Sichuan Province, ChinaDepartment of Pharmacy, Chengdu Medical College, 610500, Sichuan Province, ChinaDepartment of Basic Medical Science, Chengdu Medical College, 610500, Sichuan Province, ChinaDepartment of Basic Medical Science, Chengdu Medical College, 610500, Sichuan Province, ChinaDepartment of Public Health, Chengdu Medical College, 610500, Sichuan Province, ChinaAim: Retinal photochemical damage (RPD) plays a significant role in the development of various ocular diseases, with Caspase-1 being a key contributor. This study investigates the protective effects of Caspase-1 gene-mediated pyroptosis against RPD. Methods: Differentially expressed genes (DEGs) associated with RPD were identified through the analysis of two expression profiles from the GEO database. Correlation analysis was used to pinpoint pyroptosis-related genes (PRGs) linked to RPD. A Caspase-1 knockout 661 W cell line was generated via CRISPR-Cas9 gene editing, and single-cell colonies were screened and purified. Validation of knockout cells was performed through RT-qPCR, gene sequencing, and Western blot analysis. Comparative assays on cell proliferation, intracellular reactive oxygen species (ROS), and cytotoxicity were conducted between wild-type and Caspase-1 knockout cells under light exposure. Further RT-qPCR and Western blot experiments examined changes in the mRNA and protein levels of key pyroptosis pathway components. Results: Significant alterations in Caspase-1 expression were observed among PRGs. Homozygous Caspase-1 knockout cell lines were confirmed through RT-qPCR, genomic PCR product sequencing, and Western blot analysis. Compared to wild-type 661 W cells, Caspase-1 knockout cells exhibited higher viability and proliferation rates after 24 h of light exposure, alongside reduced LDH release. The expression of downstream pyroptosis factors at both the mRNA and protein levels was markedly decreased in the knockout group. Conclusion: CRISPR/Cas9-mediated Caspase-1 knockout enhanced the resistance of 661 W cells to photochemical damage, suggesting that Caspase-1 may serve as a potential therapeutic target for RPD-related diseases.http://www.sciencedirect.com/science/article/pii/S0890850824000434CRISPR/Cas9 SystemCaspase-1 GeneGene KnockoutPyroptosisRetinal Photochemical Damage |
| spellingShingle | Xiaoping Yu Jiayuan Peng Qian Zhong Ailin Wu Xiaoming Deng Yanfeng Zhu Caspase-1 knockout disrupts pyroptosis and protects photoreceptor cells from photochemical damage Molecular and Cellular Probes CRISPR/Cas9 System Caspase-1 Gene Gene Knockout Pyroptosis Retinal Photochemical Damage |
| title | Caspase-1 knockout disrupts pyroptosis and protects photoreceptor cells from photochemical damage |
| title_full | Caspase-1 knockout disrupts pyroptosis and protects photoreceptor cells from photochemical damage |
| title_fullStr | Caspase-1 knockout disrupts pyroptosis and protects photoreceptor cells from photochemical damage |
| title_full_unstemmed | Caspase-1 knockout disrupts pyroptosis and protects photoreceptor cells from photochemical damage |
| title_short | Caspase-1 knockout disrupts pyroptosis and protects photoreceptor cells from photochemical damage |
| title_sort | caspase 1 knockout disrupts pyroptosis and protects photoreceptor cells from photochemical damage |
| topic | CRISPR/Cas9 System Caspase-1 Gene Gene Knockout Pyroptosis Retinal Photochemical Damage |
| url | http://www.sciencedirect.com/science/article/pii/S0890850824000434 |
| work_keys_str_mv | AT xiaopingyu caspase1knockoutdisruptspyroptosisandprotectsphotoreceptorcellsfromphotochemicaldamage AT jiayuanpeng caspase1knockoutdisruptspyroptosisandprotectsphotoreceptorcellsfromphotochemicaldamage AT qianzhong caspase1knockoutdisruptspyroptosisandprotectsphotoreceptorcellsfromphotochemicaldamage AT ailinwu caspase1knockoutdisruptspyroptosisandprotectsphotoreceptorcellsfromphotochemicaldamage AT xiaomingdeng caspase1knockoutdisruptspyroptosisandprotectsphotoreceptorcellsfromphotochemicaldamage AT yanfengzhu caspase1knockoutdisruptspyroptosisandprotectsphotoreceptorcellsfromphotochemicaldamage |