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.
ACTA2 Double Nickase Plasmid (h): sc-400003-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ACTA2 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
- ACTA2 Double Nickase Plasmid (h) and ACTA2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ACTA2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: α-Actin Antibody (1A4): sc-32251
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ACTA2 Double Nickase Plasmid (h) | sc-400003-NIC | 20 µg | $410.00 | |||
ACTA2 Double Nickase Plasmid (h2) | sc-400003-NIC-2 | 20 µg | $410.00 |
ACTA2 encodes α-smooth muscle actin (ACTA2), a core contractile filament protein enriched in vascular smooth muscle cells and myofibroblasts. By polymerizing into F-actin stress fibers and engaging myosin motors, ACTA2 supports cytoskeletal tension, focal adhesion dynamics, and mechanotransduction programs that shape cell migration and extracellular matrix remodeling. ACTA2 activity intersects with RhoA/ROCK signaling, TGF-β–driven differentiation, and SRF/MRTF transcriptional control of contractile gene expression. Dysregulation or mutation of ACTA2 is implicated in vascular smooth muscle dysfunction and remodeling phenotypes, including thoracic aortic aneurysm/dissection risk, and is widely studied in contexts of fibrosis and abnormal contractility.
ACTA2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ACTA2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ACTA2. 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 ACTA2 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 ACTA2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.