AIP4 Double Nickase Plasmid (m): sc-421158-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- AIP4 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
- AIP4 Double Nickase Plasmid (m) and AIP4 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Itch. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
AIP4 Double Nickase Plasmid (m) | sc-421158-NIC | 20 µg | $410.00 |
Mouse Itch encodes AIP4, a HECT-type E3 ubiquitin ligase that catalyzes ubiquitin transfer to regulate protein turnover, trafficking, and signaling amplitude. AIP4 modulates immune and inflammatory pathways including T cell activation and differentiation, and it interfaces with receptor-proximal signaling networks such as Notch and NF-κB through ubiquitination-dependent control of key adaptors and receptors. By shaping ubiquitin-dependent sorting and degradation, ITCH influences apoptosis, cellular stress responses, and maintenance of immune homeostasis. Dysregulated ITCH/AIP4 activity has been linked to immune dysregulation phenotypes and altered signaling outputs relevant to inflammatory and autoimmune-like processes in experimental models.
AIP4 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Itch locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Itch. 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 Itch 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 Itch-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.