COL7A1 Double Nickase Plasmid (h): sc-401235-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- COL7A1 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
- COL7A1 Double Nickase Plasmid (h) and COL7A1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting COL7A1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: COL7A1 Antibody (4D2): sc-33710
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
COL7A1 Double Nickase Plasmid (h) | sc-401235-NIC | 20 µg | $410.00 | |||
COL7A1 Double Nickase Plasmid (h2) | sc-401235-NIC-2 | 20 µg | $410.00 |
COL7A1 encodes type VII collagen (COL7A1), a key structural component of anchoring fibrils that secure the epidermal basement membrane to the underlying dermis and support skin integrity. This extracellular matrix protein contributes to basement membrane assembly and stability through interactions with laminins, type IV collagen, and other dermal–epidermal junction proteins, linking matrix organization to cell–matrix adhesion and mechanotransduction. Disruption of COL7A1 function is strongly associated with inherited skin fragility phenotypes, including dystrophic epidermolysis bullosa, making it a widely studied locus for epithelial biology and matrix remodeling. COL7A1 is also used as a molecular entry point to investigate wound repair processes, keratinocyte–fibroblast crosstalk, and extracellular matrix dynamics in human cell models.
COL7A1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the COL7A1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within COL7A1. 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 COL7A1 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 COL7A1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.