ASK 1 Double Nickase Plasmid (m): sc-424043-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ASK 1 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
- ASK 1 Double Nickase Plasmid (m) and ASK 1 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Map3k5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ASK 1 Antibody (H-2): sc-390275
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ASK 1 Double Nickase Plasmid (m) | sc-424043-NIC | 20 µg | $410.00 | |||
ASK 1 Double Nickase Plasmid (m2) | sc-424043-NIC-2 | 20 µg | $410.00 |
Mouse Map3k5 encodes apoptosis signal-regulating kinase 1 (ASK1), a MAP3K that couples oxidative stress, endoplasmic reticulum stress, and inflammatory cues to downstream JNK and p38 MAPK signaling. ASK1 is activated by dissociation from redox regulators such as thioredoxin and by TRAF-mediated inputs, promoting phosphorylation cascades that influence apoptosis, cytokine production, and cellular stress adaptation. Through these pathways, ASK1 helps shape innate immune responses and stress-induced cell fate decisions in diverse tissues. Dysregulated ASK1 signaling has been linked to pathological inflammation and tissue injury processes, making it a common target in mechanistic studies of stress signaling networks.
ASK 1 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Map3k5 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Map3k5. 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 Map3k5 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 Map3k5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.