Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
IRAP Double Nickase Plasmid (h): sc-403731-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- IRAP 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
- IRAP Double Nickase Plasmid (h) and IRAP Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting LNPEP. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: IRAP Antibody (F-5): sc-365300
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
IRAP Double Nickase Plasmid (h) | sc-403731-NIC | 20 µg | $410.00 | |||
IRAP Double Nickase Plasmid (h2) | sc-403731-NIC-2 | 20 µg | $410.00 |
Leucyl/cystinyl aminopeptidase (LNPEP), also known as insulin-responsive aminopeptidase (IRAP), is a zinc-dependent M1 metallopeptidase that localizes to endosomal compartments and participates in regulated membrane trafficking. In insulin-responsive cells, IRAP co-traffics with GLUT4 storage vesicles and is mobilized to the plasma membrane following PI3K–AKT signaling, linking LNPEP to glucose uptake and vesicle recycling processes. IRAP also contributes to antigen cross-presentation through endosomal peptide trimming, shaping MHC class I peptide repertoires and influencing immune surveillance. Dysregulated LNPEP/IRAP activity or trafficking has been associated with altered metabolic homeostasis and inflammatory signaling contexts, making it relevant for studies of insulin action, endosomal proteolysis, and immune-related pathways.
IRAP Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the LNPEP locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within LNPEP. 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 LNPEP 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 LNPEP-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.