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.
PLAP Double Nickase Plasmid (h): sc-400629-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PLAP 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
- PLAP Double Nickase Plasmid (h) and PLAP Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ALPP. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PLAP Antibody (8B6): sc-47691
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PLAP Double Nickase Plasmid (h) | sc-400629-NIC | 20 µg | $410.00 | |||
PLAP Double Nickase Plasmid (h2) | sc-400629-NIC-2 | 20 µg | $410.00 |
Human ALPP encodes placental alkaline phosphatase (PLAP), a glycosylphosphatidylinositol (GPI)-anchored ectoenzyme on the cell surface that hydrolyzes phosphate monoesters and can influence extracellular nucleotide and phosphate availability. PLAP activity interfaces with membrane microdomain organization and contributes to processes such as cell differentiation, adhesion, and signaling at the plasma membrane. ALPP is physiologically prominent in placental tissue and is also used as a molecular marker in studies of ectopic expression and lineage state changes in transformed and germ cell–associated contexts. Altered ALPP/PLAP expression patterns are frequently leveraged to interrogate pathways linked to developmental regulation, membrane trafficking, and cell-state transitions.
PLAP Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ALPP locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ALPP. 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 ALPP 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 ALPP-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.