1. Enhancing Temozolomide (TMZ) chemosensitivity using CRISPR-dCas9-mediated downregulation of O 6 -methylguanine DNA methyltransferase (MGMT).
- Author
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Yousefi Y, Nejati R, Eslahi A, Alizadeh F, Farrokhi S, Asoodeh A, and Mojarrad M
- Subjects
- Humans, DNA Repair Enzymes genetics, DNA Repair Enzymes metabolism, Brain Neoplasms genetics, Brain Neoplasms drug therapy, Brain Neoplasms pathology, DNA Modification Methylases genetics, DNA Modification Methylases metabolism, Tumor Suppressor Proteins genetics, Tumor Suppressor Proteins metabolism, HEK293 Cells, Drug Resistance, Neoplasm genetics, DNA Methylation drug effects, Gene Expression Regulation, Neoplastic drug effects, Cell Line, Tumor, Promoter Regions, Genetic, Temozolomide pharmacology, Antineoplastic Agents, Alkylating pharmacology, Antineoplastic Agents, Alkylating therapeutic use, CRISPR-Cas Systems, Down-Regulation, Glioblastoma genetics, Glioblastoma drug therapy
- Abstract
Purpose: Glioblastoma (GBM) stands out as the most prevalent and aggressive intracranial tumor, notorious for its poor prognosis. The current standard-of-care for GBM patients involves surgical resection followed by radiotherapy, combined with concurrent and adjuvant chemotherapy using Temozolomide (TMZ). The effectiveness of TMZ primarily relies on the activity of O
6 -methylguanine DNA methyltransferase (MGMT), which removes alkyl adducts from the O6 position of guanine at the DNA level, thereby counteracting the toxic effects of TMZ., Method: In this study, we employed fusions of catalytically-inactive Cas9 (dCas9) to DNA methyltransferases (dCas9-DNMT3A) to selectively downregulation MGMT transcription by inducing methylation at MGMT promoter and K-M enhancer., Result: Our findings demonstrate a significant reduction in MGMT expression, leading to intensified TMZ sensitivity in the HEK293T cell line., Conclusion: This study serves as a proof of concept for the utilization of CRISPR-based gene suppression to overcome TMZ resistance and enhance the lethal effect of TMZ in glioblastoma tumor cells., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)- Published
- 2024
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