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.
Fhit Double Nickase Plasmid (h): sc-404220-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Fhit 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
- Fhit Double Nickase Plasmid (h) and Fhit Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting FHIT. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Fhit Antibody (G-4): sc-390481
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Fhit Double Nickase Plasmid (h) | sc-404220-NIC | 20 µg | $410.00 | |||
Fhit Double Nickase Plasmid (h2) | sc-404220-NIC-2 | 20 µg | $410.00 |
FHIT encodes Fhit, a histidine triad (HIT) family nucleotide-binding protein frequently implicated in maintenance of genomic stability and cellular homeostasis. Fhit participates in stress-responsive signaling and is linked to regulation of proliferation and apoptosis, with functional connections to DNA damage responses and checkpoint control. Altered FHIT expression or disruption at the common fragile site FRA3B is observed across multiple tumor types, making it a widely used model for studying tumor suppressor loss and chromosomal fragility. In cell-based systems, FHIT perturbation is used to examine how genome instability and aberrant growth signaling emerge from loss of a conserved regulatory node.
Fhit Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the FHIT locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within FHIT. 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 FHIT 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 FHIT-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.