Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
MGMT Double Nickase Plasmid (h): sc-400720-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MGMT Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- MGMT Double Nickase Plasmid (h) and MGMT Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MGMT. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: MGMT Antibody (E-1): sc-166528
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MGMT Double Nickase Plasmid (h) | sc-400720-NIC | 20 µg | $410.00 | |||
MGMT Double Nickase Plasmid (h2) | sc-400720-NIC-2 | 20 µg | $410.00 |
O6-methylguanine-DNA methyltransferase (MGMT) is a direct DNA repair enzyme that removes alkyl adducts from the O6 position of guanine by transferring the lesion to an active-site cysteine in a single-use “suicide” reaction. This activity protects genomic integrity by preventing miscoding and replication-associated double-strand breaks that can arise from persistent O6-alkylguanine lesions. MGMT functions within the broader DNA damage response and repair network that interfaces with replication stress and mismatch repair–dependent processing of alkylation damage. Altered MGMT expression or promoter methylation is widely studied as a determinant of cellular responses to alkylating damage and is relevant to tumor biology, mutational signatures, and DNA repair pathway crosstalk.
MGMT Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MGMT locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MGMT. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt MGMT function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of MGMT-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.