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.
beta Actin Double Nickase Plasmid (h): sc-400000-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- beta Actin 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
- beta Actin Double Nickase Plasmid (h) and beta Actin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ACTB. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: beta Actin Antibody (C4): sc-47778
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
beta Actin Double Nickase Plasmid (h) | sc-400000-NIC | 20 µg | $410.00 | |||
beta Actin Double Nickase Plasmid (h2) | sc-400000-NIC-2 | 20 µg | $410.00 |
ACTB encodes human beta actin, a highly conserved cytoskeletal protein that polymerizes into microfilaments to support cell shape, mechanical integrity, and intracellular transport. Beta actin dynamics coordinate with actin-binding proteins in pathways controlling cell adhesion, migration, cytokinesis, and endocytosis, and couple to signaling networks such as Rho-family GTPase regulation of actin remodeling. Because actin architecture influences chromatin organization and mechanotransduction, ACTB is frequently used as a reference for cellular homeostasis while also serving as a functional node in studies of motility and cellular stress responses. Perturbations in actin regulation and ACTB-associated cytoskeletal processes are implicated in diverse disease-relevant phenotypes, including altered invasion, developmental defects, and cytoskeletal pathologies.
beta Actin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ACTB locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ACTB. 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 ACTB 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 ACTB-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.