GTBP Double Nickase Plasmid (m): sc-421718-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GTBP Double Nickase Plasmid (m) 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
- GTBP Double Nickase Plasmid (m) and GTBP Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Msh6. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GTBP Antibody (E-8): sc-137015
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GTBP Double Nickase Plasmid (m) | sc-421718-NIC | 20 µg | $410.00 | |||
GTBP Double Nickase Plasmid (m2) | sc-421718-NIC-2 | 20 µg | $410.00 |
Mouse Msh6 encodes GTBP, a core component of the MutSα mismatch recognition complex with MSH2 that detects base–base mismatches and insertion/deletion loops arising during DNA replication and recombination. By initiating DNA mismatch repair, GTBP helps maintain genome stability, suppresses mutation accumulation, and interfaces with replication-coupled repair and damage signaling processes. Disruption of Msh6 perturbs mismatch repair fidelity, elevates microsatellite instability, and alters cellular responses to replication stress. In biomedical research, Msh6/GTBP is widely used to study mutagenesis mechanisms and genotype–phenotype relationships in cancer-associated DNA repair pathways.
GTBP Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Msh6 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Msh6. 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 Msh6 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 Msh6-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.