Perlecan Double Nickase Plasmid (h): sc-400823-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Perlecan 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
- Perlecan Double Nickase Plasmid (h) and Perlecan Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HSPG2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Perlecan Antibody (E-6): sc-377219
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Perlecan Double Nickase Plasmid (h) | sc-400823-NIC | 20 µg | $410.00 | |||
Perlecan Double Nickase Plasmid (h2) | sc-400823-NIC-2 | 20 µg | $410.00 |
HSPG2 encodes perlecan, a large heparan sulfate proteoglycan of basement membranes and pericellular matrices that binds growth factors and structural ECM components to regulate tissue architecture. Perlecan modulates cell adhesion, mechanotransduction, and morphogen signaling by organizing gradients and co-receptor interactions in pathways such as FGF, VEGF, and TGF-β, influencing angiogenesis, chondrogenesis, and wound responses. Its extracellular matrix functions are tightly linked to vascular integrity and cartilage homeostasis, and altered HSPG2 expression or structure has been associated with congenital skeletal disorders and tumor microenvironment remodeling relevant to invasion and stromal signaling studies. As a matrix organizer, perlecan is frequently examined in endothelial, smooth muscle, and stromal cell models to dissect extracellular cues that control proliferation and migration.
Perlecan Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HSPG2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HSPG2. 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 HSPG2 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 HSPG2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.