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.
VPS33B Double Nickase Plasmid (h): sc-406200-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- VPS33B 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
- VPS33B Double Nickase Plasmid (h) and VPS33B Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting VPS33B. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: VPS33B Antibody (G-9): sc-398322
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
VPS33B Double Nickase Plasmid (h) | sc-406200-NIC | 20 µg | $410.00 | |||
VPS33B Double Nickase Plasmid (h2) | sc-406200-NIC-2 | 20 µg | $410.00 |
VPS33B encodes a core component of the HOPS/CORVET-class tethering machinery that coordinates endosome maturation, vesicle docking, and SNARE-dependent membrane fusion. Through regulation of endolysosomal trafficking and polarized sorting, VPS33B supports proper delivery of cargo to lysosomes and specialized secretory pathways in differentiated cells. Disrupted VPS33B function has been linked to defects in epithelial polarity, platelet granule biogenesis, and biliary and renal physiology, making it relevant to studies of multisystem cholestatic and bleeding phenotypes. Its role in vesicle-mediated transport also intersects with pathways controlling autophagy, membrane protein turnover, and organelle homeostasis.
VPS33B Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the VPS33B locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within VPS33B. 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 VPS33B 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 VPS33B-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.