COL12A1 Double Nickase Plasmid (h): sc-402361-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- COL12A1 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
- COL12A1 Double Nickase Plasmid (h) and COL12A1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting COL12A1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: COL12A1 Antibody (A-11): sc-166020
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
COL12A1 Double Nickase Plasmid (h) | sc-402361-NIC | 20 µg | $410.00 | |||
COL12A1 Double Nickase Plasmid (h2) | sc-402361-NIC-2 | 20 µg | $410.00 |
Human COL12A1 encodes collagen type XII alpha 1 chain, a fibril-associated collagen that localizes to the extracellular matrix and interfaces with collagen I–containing fibrils to regulate matrix architecture, tensile properties, and cell–matrix communication. Through its roles in collagen fibrillogenesis, tissue organization, and mechanotransduction, COL12A1 contributes to cytoskeletal coupling and signaling pathways that shape adhesion and migration programs. Altered COL12A1 expression or function has been linked to heritable connective tissue disorders and phenotypes affecting muscle and skeletal integrity, making it relevant to studies of matrix remodeling and developmental biology. In cancer and fibrosis research, COL12A1 is frequently examined as part of stromal extracellular matrix signatures associated with changes in tissue stiffness and invasive behavior.
COL12A1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the COL12A1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within COL12A1. 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 COL12A1 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 COL12A1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.