CRISPR-dCas9 as Potential therapeuty for Glioblastoma

The most common and often-fatal type of primary brain cancer is multiform Glioblastoma. Although many attempts have been made to treat this cancer, it still has a high recurrence rate and poor prognosis. MGMT is most important prognostic marker and also therapeutic outcome predictor of glioblastoma. The protein encoded by MGMT gene is a DNA repair enzyme (O6-methylguanine-DNA-methyltransferase) that protects cell genome from methylation and can abrogate the effects of alkylating chemotherapy such as temozolamide. Using CRISPR system mediated gene suppression we suppressed MGMT expression and induced Temozolamide sensitivity in U87 cells.Methods:MGMT promoter targeting sgRNA was designed using MIT online database and cloned in TRE-KRAB-dCas9-IRES-GFP plasmid. Produced plasmid was transfected into U87 cells. MGMT gene expression levels in treated and control cells was measured using Real Time PCR. Temozolamide antiproliferative effects was compared between transfected cells and untreated control cells using MTT and cell cytotoxicity test.Results:Our results showed that CRISPR system mediated gene suppression dramatically attenuates MGMT expression and also cancerous phenotype of U87 glioblastoma cells.Conclusion:According to our results CRISPR system mediated gene suppression could be used for MGMT silencing. This strategy can be used as a new adjuvant in fighting against glioblastoma.

کلیدواژه ها:

MGMT ، CRISPR-dCas9 ، Glioblastoma.