2 results about "Mgmt methylation" patented technology

The invention discloses a human MGMT methylation detection reagent based on digital PCR (Polymerase Chain Reaction). The reagent comprises an MGMT upstream primer F1 with a sequence as shown in SEQ ID NO.1, an MGMT downstream primer R1 with a sequence as shown in SEQ ID NO.2, an MGMT probe P1 with a sequence as shown in SEQ ID NO.3, an ACTB upstream primer F with a sequence as shown in SEQ ID NO.4, an ACTB downstream primer R with a sequence as shown in SEQ ID NO.5 and an ACTB probe P with a sequence as shown in SEQ ID NO.6, the MGMT probe P1 and the ACTB probe P are respectively marked with different detection groups. The detection reagent, the kit and the detection method are high in sensitivity, high in specificity and good in detection accuracy.

Malignant glioma has a poor prognosis, despite current therapeutic modalities. Standard of care therapy consists of surgical resection, fractionated radiotherapy concurrently administered with temozolomide (TMZ), a DNA-alkylating chemotherapeutic agent, followed by adjuvant cycles of TMZ. O6-methylguanine DNA methyltransferase (MGMT) is a DNA repair enzyme which removes alkylated lesions from tumor DNA, thereby promoting chemoresistance. MGMT promoter methylation is a known predictor of temozolomide responsiveness; patients with unmethylated MGMT have a poorer prognosis. We employed deactivated Cas9-CRISPR technology to effectively target methylation in the vicinity of the MGMT methylation-specific polymerase chain reaction (MSP) region, as mediated by the catalytic domain of DNA methyltransferase DNMT3A. In doing so, we discovered that the selected dCas9 / CRISPR-mediated methylation of certain regions of MGMT gene sequences can be used to enhance chemosensitivity in malignant glioma cells.