COL1A1 Double Nickase Plasmid (h): sc-400064-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- COL1A1 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
- COL1A1 Double Nickase Plasmid (h) and COL1A1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting COL1A1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: COL1A1 Antibody (H-7): sc-518272
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
COL1A1 Double Nickase Plasmid (h) | sc-400064-NIC | 20 µg | $410.00 | |||
COL1A1 Double Nickase Plasmid (h2) | sc-400064-NIC-2 | 20 µg | $410.00 |
COL1A1 encodes the pro-α1 chain of type I collagen, a major structural component of the extracellular matrix that assembles into fibrillar collagen and provides tensile strength to connective tissues. Its biosynthesis and maturation involve ER processing, proline/lysine hydroxylation, triple-helix formation, secretion, and extracellular fibrillogenesis, linking COL1A1 to ECM organization, cell–matrix adhesion, and tissue remodeling pathways. COL1A1 expression is tightly coupled to fibroblast activation programs and signaling inputs such as TGF-β that coordinate matrix deposition and wound repair. Genetic or regulatory perturbation of COL1A1 is associated with connective tissue and skeletal phenotypes and is widely studied in the context of fibrosis and tumor microenvironment remodeling.
COL1A1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the COL1A1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within COL1A1. 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 COL1A1 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 COL1A1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.