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.
γ-GCSc Double Nickase Plasmid (m): sc-420573-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- γ-GCSc 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
- γ-GCSc Double Nickase Plasmid (m) and γ-GCSc Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Gclc. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: γ-GCSc Antibody (H-5): sc-390811
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
γ-GCSc Double Nickase Plasmid (m) | sc-420573-NIC | 20 µg | $410.00 |
Gclc encodes the catalytic subunit of glutamate–cysteine ligase (γ-GCSc), the rate-limiting enzyme for de novo glutathione biosynthesis. By controlling cellular glutathione levels, γ-GCSc influences redox homeostasis, detoxification of electrophiles and xenobiotics, and maintenance of thiol-dependent signaling. Gclc activity interfaces with oxidative stress response networks, including NRF2-regulated antioxidant programs, and impacts mitochondrial function and cellular susceptibility to reactive oxygen species. Dysregulated glutathione metabolism and impaired Gclc function have been linked to oxidative damage phenotypes relevant to inflammation, neurodegeneration, metabolic stress, and toxicant sensitivity in biomedical models.
γ-GCSc Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Gclc locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Gclc. 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 Gclc 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 Gclc-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.