gp91phox/CYBB/NOX2 Double Nickase Plasmid (h): sc-400222-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- gp91phox/CYBB/NOX2 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
- gp91phox/CYBB/NOX2 Double Nickase Plasmid (h) and gp91phox/CYBB/NOX2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CYBB. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: gp91phox/CYBB/NOX2 Antibody (54.1): sc-130543
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
gp91phox/CYBB/NOX2 Double Nickase Plasmid (h) | sc-400222-NIC | 20 µg | $410.00 | |||
gp91phox/CYBB/NOX2 Double Nickase Plasmid (h2) | sc-400222-NIC-2 | 20 µg | $410.00 |
CYBB encodes gp91phox (NOX2), the catalytic membrane subunit of the phagocyte NADPH oxidase complex responsible for generating reactive oxygen species during the oxidative burst. NOX2 activity supports innate immune defense by driving superoxide production in neutrophils and macrophages and influences downstream redox-sensitive signaling pathways, including MAPK and NF-κB, that shape inflammatory responses. Disruption of CYBB function is linked to impaired microbial killing and altered inflammatory homeostasis, making it a key node in studies of phagocyte biology. CYBB is also used as a model locus to investigate assembly and regulation of multi-subunit oxidase complexes and compartmentalized ROS signaling.
gp91phox/CYBB/NOX2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CYBB locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CYBB. 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 CYBB 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 CYBB-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.