COL8A1 Double Nickase Plasmid (h): sc-405174-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- COL8A1 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
- COL8A1 Double Nickase Plasmid (h) and COL8A1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting COL8A1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
COL8A1 Double Nickase Plasmid (h) | sc-405174-NIC | 20 µg | $410.00 | |||
COL8A1 Double Nickase Plasmid (h2) | sc-405174-NIC-2 | 20 µg | $410.00 |
COL8A1 encodes the alpha 1 chain of type VIII collagen, a short-chain, non-fibrillar collagen enriched in specialized extracellular matrices such as corneal endothelium and vascular basement membrane-associated regions. COL8A1 contributes to matrix assembly, cell–matrix adhesion, and tissue biomechanics, influencing signaling crosstalk between the extracellular matrix and intracellular pathways that regulate migration, proliferation, and differentiation. Altered COL8A1 expression or matrix deposition has been linked to endothelial dysfunction and remodeling processes relevant to ocular physiology and vascular pathology, making it a useful target for studying extracellular matrix homeostasis and mechanotransduction. In vitro and in vivo models that perturb COL8A1 help interrogate matrix-driven changes in cellular behavior, barrier properties, and stress responses.
COL8A1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the COL8A1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within COL8A1. 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 COL8A1 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 COL8A1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.