VPS54 CRISPR/Cas9 KO Plasmid (m): sc-434184
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- VPS54 CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 VPS54 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: VPS54 Antibody (B-4): sc-398300
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
VPS54 CRISPR/Cas9 KO Plasmid (m) | sc-434184 | 20 µg | $397.00 |
Overview
Vps54 encodes VPS54, a core subunit of the GARP (Golgi-associated retrograde protein) tethering complex that promotes endosome-to-trans-Golgi network retrograde trafficking and supports proper Golgi organization. By coordinating vesicle capture and fusion, VPS54 helps maintain membrane protein recycling, glycosylation-dependent cargo sorting, and overall secretory pathway homeostasis. Disruption of GARP components perturbs endolysosomal transport, triggers cellular stress responses, and can alter neuronal maintenance pathways through defects in trafficking and organelle integrity. Because membrane trafficking is tightly coupled to proteostasis and signaling, Vps54 is frequently examined in studies of neurodevelopmental and neurodegeneration-relevant mechanisms.
VPS54 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Vps54 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Vps54 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 Vps54 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 VPS54 protein expression.
This CRISPR knockout system enables efficient generation of Vps54-deficient cell models for investigation of VPS54 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Vps54 exon(s) critical for VPS54 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 Vps54 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by VPS54 CRISPR/Cas9 KO Plasmid (m) and VPS54 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Vps54 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by VPS54 HDR Plasmid (m) and VPS54 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Vps54 homology arms to support homology-directed repair at defined Vps54 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.