cystatin D Double Nickase Plasmid (h): sc-405049-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- cystatin D 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
- cystatin D Double Nickase Plasmid (h) and cystatin D Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CST5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: cystatin D Antibody (F-3): sc-398553
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
cystatin D Double Nickase Plasmid (h) | sc-405049-NIC | 20 µg | $410.00 | |||
cystatin D Double Nickase Plasmid (h2) | sc-405049-NIC-2 | 20 µg | $410.00 |
CST5 encodes cystatin D, a secreted type 2 cystatin that inhibits lysosomal and extracellular cysteine proteases such as cathepsins, thereby helping regulate proteolysis, antigen processing, and epithelial barrier homeostasis. By modulating cathepsin activity, cystatin D can influence extracellular matrix turnover and protease-dependent signaling pathways relevant to tissue remodeling and inflammatory responses. Altered CST5 expression or protease–inhibitor imbalance has been associated with changes in tumor microenvironment behavior, invasion-related proteolysis, and mucosal pathophysiology, supporting its use as a mechanistic node in studies of proteostasis and epithelial biology. CST5 is therefore a useful target for dissecting how cathepsin control shapes cellular differentiation, stress responses, and intercellular communication in human model systems.
cystatin D Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CST5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CST5. 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 CST5 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 CST5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.