CRISPR/Cas9-Mediated genome editing in grapevine protoplasts; [Asma protoplastlarında CRISPR/Cas9 aracılı genom düzenleme]
| dc.contributor.author | Kaya H.B. | |
| dc.date.accessioned | 2025-04-10T11:01:39Z | |
| dc.date.available | 2025-04-10T11:01:39Z | |
| dc.date.issued | 2025 | |
| dc.description.abstract | Objective: This study aims to perform targeted mutation in grapevine protoplasts using the CRISPR/Cas9-mediated genome editing method. Material and Methods: For CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9)-mediated genome editing in the Chardonnay cultivar, a gRNA design targeting the desired gene was performed, resulting in obtaining a final CRISPR/Cas9 vector containing both the gRNA and Cas9 and GFP genes. Protoplast isolation and transformation were performed using leaves, followed by analysis of transformation and mutation efficiency. Results: In the study, Chardonnay leaf protoplast isolation produced 1x107 protoplasts per 1 g of fresh leaves. The vector targeting the VvPDS gene (~10 kb) achieved a transformation efficiency of 40-60%, while the vector containing only the GFP gene (~3 kb) reached 80-90% efficiency. Vector size notably impacted transformation, with larger vectors reducing efficiency. Despite successful transformation, the presence of the targeted mutation could not be confirmed. Conclusion: The study successfully completed all stages from gRNA design, the initial step of CRISPR/Cas9-mediated genome editing in protoplasts, to the final protoplast transformation stage, showcasing the system's seamless usability. The protocols applied and the results obtained can be utilized in future studies aimed at implementing targeted mutations in grapevines. © 2025 Ege Universitesi. All rights reserved. | |
| dc.identifier.DOI-ID | 10.20289/zfdergi.1432614 | |
| dc.identifier.uri | http://hdl.handle.net/20.500.14701/43596 | |
| dc.publisher | Ege Universitesi | |
| dc.title | CRISPR/Cas9-Mediated genome editing in grapevine protoplasts; [Asma protoplastlarında CRISPR/Cas9 aracılı genom düzenleme] | |
| dc.type | Article |