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.
TNAP Double Nickase Plasmid (h): sc-400784-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TNAP 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
- TNAP Double Nickase Plasmid (h) and TNAP Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ALPL. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TNAP Antibody (TRA-2-39): sc-21708
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TNAP Double Nickase Plasmid (h) | sc-400784-NIC | 20 µg | $410.00 | |||
TNAP Double Nickase Plasmid (h2) | sc-400784-NIC-2 | 20 µg | $410.00 |
ALPL encodes tissue-nonspecific alkaline phosphatase (TNAP), a glycosylphosphatidylinositol-anchored ectoenzyme that hydrolyzes extracellular phosphate monoesters to regulate inorganic phosphate and pyrophosphate balance. By degrading pyrophosphate, TNAP modulates mineralization processes and supports proper bone and tooth matrix maturation, while also influencing purinergic signaling through nucleotide dephosphorylation in the extracellular milieu. ALPL/TNAP activity intersects with pathways controlling phosphate homeostasis and extracellular matrix calcification, and altered function is linked to hypophosphatasia and pathological calcification phenotypes. As a surface-exposed enzyme with measurable catalytic output, TNAP provides a tractable node for studying genotype-to-phenotype relationships in mineral metabolism and cell differentiation models.
TNAP Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ALPL locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ALPL. 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 ALPL 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 ALPL-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.