Cystinosin Double Nickase Plasmid (h): sc-405040-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Cystinosin 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
- Cystinosin Double Nickase Plasmid (h) and Cystinosin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CTNS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Cystinosin Antibody (F-12): sc-100703
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Cystinosin Double Nickase Plasmid (h) | sc-405040-NIC | 20 µg | $410.00 | |||
Cystinosin Double Nickase Plasmid (h2) | sc-405040-NIC-2 | 20 µg | $410.00 |
CTNS encodes cystinosin, a lysosomal membrane H+-driven cystine transporter that mediates cystine efflux from the lysosomal lumen to the cytosol, supporting intracellular thiol homeostasis. By controlling cystine handling within the endolysosomal system, cystinosin influences lysosomal function, redox balance, and downstream cellular stress responses. Disruption of CTNS perturbs lysosomal amino acid transport and can alter autophagy-lysosome pathway dynamics, making it a key node for studying organelle proteostasis. CTNS loss-of-function is associated with cystinosis, a lysosomal storage disorder characterized by intracellular cystine accumulation and multisystem cellular dysfunction.
Cystinosin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CTNS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CTNS. 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 CTNS 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 CTNS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.