GCK Double Nickase Plasmid (h): sc-400852-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GCK 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
- GCK Double Nickase Plasmid (h) and GCK Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GCK. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GCK Antibody (G-6): sc-17819
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GCK Double Nickase Plasmid (h) | sc-400852-NIC | 20 µg | $410.00 | |||
GCK Double Nickase Plasmid (h2) | sc-400852-NIC-2 | 20 µg | $410.00 |
Glucokinase (GCK) encodes the hexokinase IV enzyme that phosphorylates glucose to glucose-6-phosphate, functioning as a key glucose sensor that couples extracellular glucose availability to cellular metabolism. In pancreatic β cells, GCK activity helps set the threshold for glucose-stimulated insulin secretion, while in hepatocytes it supports glycolytic flux and glycogen synthesis through integration with glycolysis, gluconeogenesis regulation, and carbohydrate-responsive metabolic programs. By controlling glucose entry into central carbon metabolism, GCK influences ATP/ADP balance, metabolite signaling, and downstream pathways linked to insulin secretion and hepatic glucose utilization. Genetic and functional perturbations of GCK are strongly associated with altered glucose homeostasis and monogenic forms of dysglycemia, making it a widely used target for mechanistic studies of metabolic regulation.
GCK Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GCK locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GCK. 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 GCK 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 GCK-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.