Nrf2 Double Nickase Plasmid (h): sc-400017-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Nrf2 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
- Nrf2 Double Nickase Plasmid (h) and Nrf2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting NFE2L2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Nrf2 Antibody (H-6): sc-518033
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Nrf2 Double Nickase Plasmid (h) | sc-400017-NIC | 20 µg | $410.00 | |||
Nrf2 Double Nickase Plasmid (h2) | sc-400017-NIC-2 | 20 µg | $410.00 |
NFE2L2 encodes nuclear factor erythroid 2–related factor 2 (Nrf2), a basic leucine zipper transcription factor that coordinates cellular defense against oxidative and electrophilic stress. Upon stabilization and nuclear translocation, Nrf2 binds antioxidant response elements to induce genes involved in glutathione synthesis, xenobiotic detoxification, NADPH regeneration, and proteostasis, integrating redox control with metabolism and inflammatory signaling. This pathway intersects with KEAP1-dependent ubiquitination, MAPK signaling, and autophagy regulation, shaping responses to mitochondrial dysfunction and environmental toxicants. Dysregulated Nrf2 activity has been linked to altered stress tolerance and metabolic rewiring in diverse disease-relevant contexts, including carcinogenesis, neurodegeneration, and chronic inflammatory states.
Nrf2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the NFE2L2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within NFE2L2. 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 NFE2L2 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 NFE2L2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.