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.
Mel-CAM Double Nickase Plasmid (h): sc-401020-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Mel-CAM 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
- Mel-CAM Double Nickase Plasmid (h) and Mel-CAM Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting MCAM. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Mel-CAM Antibody (P1H12): sc-18837
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Mel-CAM Double Nickase Plasmid (h) | sc-401020-NIC | 20 µg | $410.00 | |||
Mel-CAM Double Nickase Plasmid (h2) | sc-401020-NIC-2 | 20 µg | $410.00 |
MCAM encodes Mel-CAM (CD146), a cell-surface immunoglobulin superfamily adhesion molecule expressed on endothelial cells and subsets of immune and stromal cells. Mel-CAM participates in cell–cell and cell–matrix interactions that influence cytoskeletal remodeling, migration, and junctional organization, with downstream effects on vascular biology and inflammatory trafficking. MCAM has been implicated in processes linked to angiogenesis and epithelial–mesenchymal-like phenotypic changes, integrating signals from the extracellular environment to regulate motility and invasion-associated programs. Dysregulated MCAM expression is frequently studied in tumor progression and metastasis models, and it is also relevant to vascular dysfunction and immune-mediated pathobiology.
Mel-CAM Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the MCAM locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within MCAM. 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 MCAM 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 MCAM-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.