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.
VPS35 Double Nickase Plasmid (h): sc-402019-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- VPS35 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
- VPS35 Double Nickase Plasmid (h) and VPS35 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting VPS35. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: VPS35 Antibody (B-5): sc-374372
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
VPS35 Double Nickase Plasmid (h) | sc-402019-NIC | 20 µg | $410.00 | |||
VPS35 Double Nickase Plasmid (h2) | sc-402019-NIC-2 | 20 µg | $410.00 |
VPS35 encodes a core component of the retromer cargo-selective complex that orchestrates endosome-to-Golgi retrieval and endosomal recycling of transmembrane proteins. By coupling to sorting nexins and WASH-dependent actin remodeling, VPS35 helps regulate receptor trafficking, lysosomal homeostasis, and the balance between recycling and degradation pathways. Disruption of VPS35 function perturbs endolysosomal dynamics and proteostasis, affecting membrane protein composition at the cell surface and within the secretory pathway. Genetic and functional studies have linked altered VPS35 activity to neurodegenerative mechanisms, particularly pathways converging on synaptic maintenance, mitochondrial quality control, and protein aggregation susceptibility.
VPS35 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the VPS35 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within VPS35. 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 VPS35 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 VPS35-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.