IAP Double Nickase Plasmid (h): sc-401544-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- IAP 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
- IAP Double Nickase Plasmid (h) and IAP Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ALPI. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: IAP Antibody (AAP.1): sc-56917
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
IAP Double Nickase Plasmid (h) | sc-401544-NIC | 20 µg | $410.00 | |||
IAP Double Nickase Plasmid (h2) | sc-401544-NIC-2 | 20 µg | $410.00 |
Human ALPI encodes intestinal alkaline phosphatase (IAP), a brush-border ectoenzyme that dephosphorylates luminal substrates and contributes to epithelial barrier homeostasis. IAP activity intersects with mucosal innate immune regulation by detoxifying pro-inflammatory phosphorylated microbial products and shaping host–microbe interactions, thereby influencing downstream inflammatory signaling in the gut. ALPI expression is tightly linked to enterocyte differentiation and nutrient handling, and altered IAP activity has been associated with intestinal inflammation and dysbiosis-related phenotypes in experimental systems. As a functional marker of small intestinal epithelium, ALPI is frequently used to interrogate pathways governing barrier integrity, microbial sensing, and epithelial maturation.
IAP Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ALPI locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ALPI. 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 ALPI 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 ALPI-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.