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.
Calpastatin Double Nickase Plasmid (h): sc-401502-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Calpastatin 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
- Calpastatin Double Nickase Plasmid (h) and Calpastatin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CAST. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Calpastatin Antibody (A-1): sc-376547
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Calpastatin Double Nickase Plasmid (h) | sc-401502-NIC | 20 µg | $410.00 | |||
Calpastatin Double Nickase Plasmid (h2) | sc-401502-NIC-2 | 20 µg | $410.00 |
Human CAST encodes calpastatin, the endogenous and highly specific inhibitor of calcium-dependent cysteine proteases calpain-1 and calpain-2. By restraining calpain-mediated limited proteolysis, calpastatin helps regulate cytoskeletal remodeling, focal adhesion turnover, membrane trafficking, and signal transduction events linked to Ca2+ dynamics and phosphorylation cascades. CAST activity influences proteostasis and cell survival programs by modulating cleavage of substrates involved in apoptosis, inflammation, and neuronal and muscle homeostasis. Dysregulated calpain–calpastatin balance has been associated with pathways relevant to neurodegeneration, cardiovascular injury responses, and cancer cell invasion, supporting CAST as a mechanistic node for studying Ca2+-regulated proteolysis.
Calpastatin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CAST locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CAST. 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 CAST 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 CAST-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.