GFAT1 Double Nickase Plasmid (h): sc-403022-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GFAT1 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
- GFAT1 Double Nickase Plasmid (h) and GFAT1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GFPT1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: GFAT1 Antibody (D-9): sc-377479
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GFAT1 Double Nickase Plasmid (h) | sc-403022-NIC | 20 µg | $410.00 | |||
GFAT1 Double Nickase Plasmid (h2) | sc-403022-NIC-2 | 20 µg | $410.00 |
GFPT1 encodes glutamine—fructose-6-phosphate amidotransferase 1 (GFAT1), the rate-limiting enzyme of the hexosamine biosynthetic pathway that converts fructose-6-phosphate and glutamine to glucosamine-6-phosphate. By controlling flux toward UDP-GlcNAc production, GFAT1 influences protein N- and O-GlcNAcylation, ER protein quality control, and metabolic sensing linked to glucose and amino acid availability. This pathway intersects with insulin signaling, stress responses, and glycoprotein synthesis, positioning GFPT1 as a key node connecting nutrient status to post-translational modification and cellular homeostasis. Altered GFPT1 activity and hexosamine pathway output have been associated with congenital myasthenic syndrome and with dysregulated glycosylation and metabolic phenotypes in human disease-relevant contexts.
GFAT1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GFPT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GFPT1. 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 GFPT1 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 GFPT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.