ACTA1 Double Nickase Plasmid (h): sc-400005-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ACTA1 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
- ACTA1 Double Nickase Plasmid (h) and ACTA1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ACTA1. 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 (733H2P): sc-517660
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ACTA1 Double Nickase Plasmid (h) | sc-400005-NIC | 20 µg | $410.00 | |||
ACTA1 Double Nickase Plasmid (h2) | sc-400005-NIC-2 | 20 µg | $410.00 |
ACTA1 encodes alpha skeletal muscle actin, a major component of the sarcomeric thin filament that polymerizes into F-actin to support myofibril assembly, force generation, and mechanotransduction. ACTA1-driven actin dynamics interface with actin–myosin cross-bridge cycling, Z-disc organization, and cytoskeletal remodeling pathways that govern muscle fiber structure and contractile performance. Pathogenic variation or dysregulated expression of ACTA1 is linked to inherited skeletal muscle disorders, including congenital myopathies, where altered filament stability and contractile mechanics contribute to cellular dysfunction. As a marker and functional node in muscle biology, ACTA1 is widely studied in myogenesis models, sarcomere biogenesis, and cytoskeletal stress responses.
ACTA1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ACTA1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ACTA1. 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 ACTA1 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 ACTA1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.