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 CRISPR/Cas9 KO Plasmid (h): sc-406200
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- VPS33B CRISPR/Cas9 Knockout (KO) Plasmid (h) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the VPS33B genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
- 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 CRISPR/Cas9 KO Plasmid (h) | sc-406200 | 20 µg | $397.00 |
Overview
VPS33B encodes a core component of the class C/VPS tethering machinery that coordinates vesicle tethering and SNARE-dependent fusion within the endolysosomal system. It functions with VPS16 and related partners to regulate endosome maturation, lysosome biogenesis, and polarized membrane trafficking, thereby influencing protein sorting, recycling, and degradative flux. Disruption of VPS33B perturbs epithelial polarity and secretory pathway organization and has been linked to multisystem trafficking disorders, including ARC syndrome, making it relevant to studies of membrane dynamics and organelle homeostasis. In immune and barrier tissues, altered VPS33B activity can impact granule/vesicle handling and membrane protein composition, connecting it to broader questions in inflammation and tissue integrity.
VPS33B CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the VPS33B gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the VPS33B together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the VPS33B open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish VPS33B protein expression.
This CRISPR knockout system enables efficient generation of VPS33B-deficient cell models for investigation of VPS33B signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting VPS33B exon(s) critical for VPS33B function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple VPS33B genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by VPS33B CRISPR/Cas9 KO Plasmid (h) and VPS33B CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the VPS33B locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by VPS33B HDR Plasmid (h) and VPS33B HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by VPS33B homology arms to support homology-directed repair at defined VPS33B target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.