Keap1 Double Nickase Plasmid (h): sc-400190-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Keap1 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
- Keap1 Double Nickase Plasmid (h) and Keap1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting KEAP1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Keap1 Antibody (G-2): sc-365626
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Keap1 Double Nickase Plasmid (h) | sc-400190-NIC | 20 µg | $410.00 | |||
Keap1 Double Nickase Plasmid (h2) | sc-400190-NIC-2 | 20 µg | $410.00 |
KEAP1 encodes Keap1, a cytosolic substrate adaptor for the CUL3–RBX1 E3 ubiquitin ligase complex that controls proteasomal turnover of NRF2 (NFE2L2) and thereby tunes the antioxidant and electrophile stress response. By sensing reactive cysteine modifications, Keap1 links redox homeostasis to transcriptional programs governing detoxification, glutathione metabolism, and xenobiotic defense, while also intersecting with autophagy via p62/SQSTM1-dependent regulation. Perturbation of the KEAP1–NRF2 axis reshapes cellular metabolism and inflammatory signaling, with broad relevance to oxidative stress biology and oncogenic stress adaptation. Dysregulated KEAP1 function is frequently studied in the context of altered NRF2 target gene expression, mitochondrial function, and resistance to environmental or metabolic stressors in human disease models.
Keap1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the KEAP1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within KEAP1. 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 KEAP1 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 KEAP1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.