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.
FX Double Nickase Plasmid (h): sc-408777-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- FX 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
- FX Double Nickase Plasmid (h) and FX Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TSTA3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: FX Antibody (43.1): sc-100531
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
FX Double Nickase Plasmid (h) | sc-408777-NIC | 20 µg | $410.00 | |||
FX Double Nickase Plasmid (h2) | sc-408777-NIC-2 | 20 µg | $410.00 |
TSTA3 encodes GDP-L-fucose synthase (FX), a cytosolic enzyme that catalyzes the final steps of the de novo GDP-fucose biosynthetic pathway from GDP-mannose. By controlling intracellular GDP-fucose availability, FX supports fucosylation of glycans on cell-surface and secreted proteins, influencing protein folding, receptor–ligand interactions, and extracellular matrix communication. Altered fucosylation is linked to changes in cell adhesion and signaling, with relevance to immune modulation, inflammation, and tumor-associated glycan remodeling. Disruption or dysregulation of TSTA3 can therefore affect glycosylation-dependent pathways that shape development and disease-associated cellular phenotypes.
FX Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TSTA3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TSTA3. 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 TSTA3 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 TSTA3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.