calsequestrin 1 Double Nickase Plasmid (h): sc-403375-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- calsequestrin 1 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
- calsequestrin 1 Double Nickase Plasmid (h) and calsequestrin 1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CASQ1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: calsequestrin 1 Antibody (D-10): sc-137080
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
calsequestrin 1 Double Nickase Plasmid (h) | sc-403375-NIC | 20 µg | $410.00 | |||
calsequestrin 1 Double Nickase Plasmid (h2) | sc-403375-NIC-2 | 20 µg | $410.00 |
CASQ1 encodes calsequestrin 1, a high-capacity Ca²⁺-binding protein localized to the sarcoplasmic reticulum lumen in skeletal muscle, where it buffers Ca²⁺ and helps shape excitation–contraction coupling. By modulating luminal Ca²⁺ availability and ryanodine receptor (RYR1) Ca²⁺ release kinetics, calsequestrin 1 contributes to Ca²⁺ homeostasis and coordinated muscle contraction/relaxation cycles. Altered CASQ1 function or expression can perturb intracellular Ca²⁺ signaling and stress responses linked to abnormal Ca²⁺ release, impaired contractility, and susceptibility to muscle dysfunction phenotypes. As a central node in sarcoplasmic reticulum Ca²⁺ handling, CASQ1 is relevant for mechanistic studies of Ca²⁺-dependent signaling, proteostasis, and metabolic remodeling in myofibers.
calsequestrin 1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CASQ1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CASQ1. 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 CASQ1 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 CASQ1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.