Calpain 5 Double Nickase Plasmid (h): sc-403881-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Calpain 5 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
- Calpain 5 Double Nickase Plasmid (h) and Calpain 5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CAPN5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Calpain 5 Antibody (A-5): sc-271271
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Calpain 5 Double Nickase Plasmid (h) | sc-403881-NIC | 20 µg | $410.00 | |||
Calpain 5 Double Nickase Plasmid (h2) | sc-403881-NIC-2 | 20 µg | $410.00 |
CAPN5 encodes calpain 5, a calcium-dependent cysteine protease in the calpain family that modulates regulated proteolysis of intracellular substrates. Calpain activity contributes to cytoskeletal remodeling, membrane dynamics, and signal transduction by coupling calcium flux to protease-dependent processing events that influence cell adhesion, migration, and survival programs. CAPN5 has been linked to retinal biology and inflammatory signaling, with genetic variation associated with inherited vitreoretinopathy and abnormal immune-like responses in ocular tissues. These features make CAPN5 a useful node for studying calcium-triggered proteostasis, stress responses, and tissue-specific degenerative mechanisms in human cell models.
Calpain 5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CAPN5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CAPN5. 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 CAPN5 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 CAPN5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.