Keap1 Double Nickase Plasmid (m): sc-424513-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Keap1 Double Nickase Plasmid (m) 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 (m) and Keap1 Double Nickase Plasmid (m2) 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 (F-10): sc-514914
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Keap1 Double Nickase Plasmid (m) | sc-424513-NIC | 20 µg | $410.00 | |||
Keap1 Double Nickase Plasmid (m2) | sc-424513-NIC-2 | 20 µg | $410.00 |
Keap1 (Kelch-like ECH-associated protein 1) is a cytosolic adaptor for a CUL3-based E3 ubiquitin ligase complex that binds NFE2L2/NRF2 and promotes its ubiquitination and proteasomal turnover under basal conditions. Oxidative or electrophilic stress modifies key cysteine residues in KEAP1, weakening NRF2 repression and enabling transcription of antioxidant, detoxification, and metabolic genes through the ARE program. This KEAP1–NRF2 axis coordinates redox homeostasis, glutathione metabolism, and xenobiotic response pathways in mouse cells, influencing inflammation, mitochondrial function, and proteostasis. Dysregulated Keap1 activity is broadly used as a mechanistic handle in models of oxidative stress biology and conditions characterized by altered redox signaling.
Keap1 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Keap1 locus in mouse 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.