V-ATPase D1 CRISPR/Cas9 KO Plasmid (m): sc-419255
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- V-ATPase D1 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 V-ATPase D1 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: V-ATPase D1 Antibody (D-4): sc-393322
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
V-ATPase D1 CRISPR/Cas9 KO Plasmid (m) | sc-419255 | 20 µg | $397.00 |
Overview
Atp6v0d1 encodes the D1 subunit of the vacuolar H\+-ATPase (V-ATPase) V0 domain, a proton pump that drives acidification of endosomes, lysosomes, and other intracellular compartments in mouse cells. V-ATPase-dependent pH regulation supports receptor-mediated endocytosis, autophagic flux, lysosomal proteolysis, and membrane trafficking, and it also contributes to organelle homeostasis coupled to mTORC1 nutrient sensing. Disruption of V-ATPase components is broadly linked to defects in vesicular sorting, impaired degradative capacity, and altered signaling outputs that can influence neurodegeneration, immune cell function, and cancer-associated metabolic adaptation. Atp6v0d1 is therefore a useful node for studying how organelle acidification intersects with proteostasis and stress-response pathways.
V-ATPase D1 CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Atp6v0d1 gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Atp6v0d1 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 Atp6v0d1 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 V-ATPase D1 protein expression.
This CRISPR knockout system enables efficient generation of Atp6v0d1-deficient cell models for investigation of V-ATPase D1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Atp6v0d1 exon(s) critical for V-ATPase D1 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 Atp6v0d1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by V-ATPase D1 CRISPR/Cas9 KO Plasmid (m) and V-ATPase D1 CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Atp6v0d1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by V-ATPase D1 HDR Plasmid (m) and V-ATPase D1 HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Atp6v0d1 homology arms to support homology-directed repair at defined Atp6v0d1 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.