Pilt Double Nickase Plasmid (m): sc-428717-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Pilt 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
- Pilt Double Nickase Plasmid (m) and Pilt Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Tjap1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Pilt Double Nickase Plasmid (m) | sc-428717-NIC | 20 µg | $410.00 |
Mouse Tjap1 encodes Pilt, a junction-associated factor implicated in organizing epithelial cell–cell contacts and coordinating cytoskeletal dynamics that influence barrier formation and tissue architecture. Pilt is linked to processes such as tight junction assembly, polarity maintenance, and regulation of paracellular permeability through protein complex scaffolding and signaling cross-talk. Perturbation of junctional homeostasis can impact proliferation, differentiation, and inflammatory signaling, making Tjap1 a useful locus for studying mechanisms that underlie epithelial dysfunction in disease-relevant contexts. Investigating Tjap1 supports functional mapping of junctional pathways, cell morphology programs, and stress responses in mouse models.
Pilt Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Tjap1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Tjap1. 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 Tjap1 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 Tjap1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.