COL4A3 Double Nickase Plasmid (h): sc-400903-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- COL4A3 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
- COL4A3 Double Nickase Plasmid (h) and COL4A3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting COL4A3. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
COL4A3 Double Nickase Plasmid (h) | sc-400903-NIC | 20 µg | $410.00 | |||
COL4A3 Double Nickase Plasmid (h2) | sc-400903-NIC-2 | 20 µg | $410.00 |
COL4A3 encodes the α3 chain of type IV collagen, a core structural component of basement membranes that assembles into the α3α4α5(IV) network with COL4A4 and COL4A5. This extracellular matrix scaffold supports epithelial and endothelial integrity and influences cell adhesion, migration, and mechanotransduction through integrin-mediated signaling and basement membrane remodeling pathways. In kidney glomeruli, COL4A3 is essential for glomerular basement membrane architecture and filtration barrier function, making it highly relevant to studies of inherited nephropathies and extracellular matrix dysregulation. Perturbation of COL4A3 is also used to model how basement membrane composition affects tissue stability and inflammatory or fibrotic responses.
COL4A3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the COL4A3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within COL4A3. 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 COL4A3 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 COL4A3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.