COL2A1 Double Nickase Plasmid (h): sc-400359-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- COL2A1 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
- COL2A1 Double Nickase Plasmid (h) and COL2A1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting COL2A1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: COL2A1 Antibody (M2139): sc-52658
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
COL2A1 Double Nickase Plasmid (h) | sc-400359-NIC | 20 µg | $410.00 | |||
COL2A1 Double Nickase Plasmid (h2) | sc-400359-NIC-2 | 20 µg | $410.00 |
COL2A1 encodes the α1 chain of type II collagen, a major fibrillar collagen in hyaline cartilage and the developing vertebral column. Its extracellular matrix assembly supports chondrocyte differentiation, tissue biomechanics, and collagen fibrillogenesis, integrating with proteoglycan-rich networks to maintain cartilage integrity. COL2A1 expression and processing intersect with matrix remodeling pathways, including regulation by SOX9-driven chondrogenic programs and feedback from TGF-β family signaling. Genetic variation or dysregulated COL2A1 function is associated with heritable skeletal dysplasias and osteoarthritis-relevant cartilage degeneration, making it a common target in musculoskeletal and extracellular matrix biology research.
COL2A1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the COL2A1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within COL2A1. 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 COL2A1 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 COL2A1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.