Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
COL5A1 Double Nickase Plasmid (h): sc-401579-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- COL5A1 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
- COL5A1 Double Nickase Plasmid (h) and COL5A1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting COL5A1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: COL5A1 Antibody (C-5): sc-166155
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
COL5A1 Double Nickase Plasmid (h) | sc-401579-NIC | 20 µg | $410.00 | |||
COL5A1 Double Nickase Plasmid (h2) | sc-401579-NIC-2 | 20 µg | $410.00 |
COL5A1 encodes the alpha 1 chain of type V collagen, a quantitatively minor but functionally critical fibrillar collagen that regulates collagen fibrillogenesis and controls the diameter and organization of type I/V collagen fibers in the extracellular matrix. Through its role in matrix assembly, COL5A1 influences cell–matrix signaling, tissue tensile strength, and remodeling processes that intersect with integrin-mediated adhesion and pathways governing fibroblast behavior. Altered COL5A1 function or expression is associated with heritable connective tissue disorders, including classical Ehlers–Danlos syndrome, and is frequently studied in contexts of skin and tendon mechanics, vascular integrity, and matrix dysregulation. As a structural ECM determinant, it is also relevant for investigating how collagen architecture modulates development, wound repair, and stromal contributions to disease phenotypes.
COL5A1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the COL5A1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within COL5A1. 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 COL5A1 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 COL5A1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.