Integrin α5/ITGA5/CD49e Double Nickase Plasmid (h): sc-400546-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Integrin α5/ITGA5/CD49e 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
- Integrin α5/ITGA5/CD49e Double Nickase Plasmid (h) and Integrin α5/ITGA5/CD49e Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ITGA5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Integrin α5/ITGA5/CD49e Antibody (C-9): sc-376199
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Integrin α5/ITGA5/CD49e Double Nickase Plasmid (h) | sc-400546-NIC | 20 µg | $410.00 | |||
Integrin α5/ITGA5/CD49e Double Nickase Plasmid (h2) | sc-400546-NIC-2 | 20 µg | $410.00 |
ITGA5 encodes integrin α5 (CD49e), an α subunit that heterodimerizes with β1 to form the α5β1 fibronectin receptor, a key mediator of cell–extracellular matrix adhesion and mechanotransduction. Engagement of fibronectin triggers focal adhesion assembly and signaling through FAK/SRC, PI3K–AKT, and MAPK pathways, coordinating actin cytoskeletal remodeling, migration, proliferation, and survival. ITGA5-dependent adhesion dynamics influence epithelial–mesenchymal transition, angiogenic programs, and tissue remodeling through crosstalk with Rho GTPases and growth factor receptor signaling. Dysregulated α5β1 activity is frequently studied in contexts of tumor invasion and metastasis, fibrosis, and vascular and inflammatory pathophysiology where altered matrix interactions drive aberrant cell behavior.
Integrin α5/ITGA5/CD49e Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ITGA5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ITGA5. 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 ITGA5 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 ITGA5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.