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.
V-ATPase A1 CRISPR/Cas9 KO Plasmid (h): sc-400975
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- V-ATPase A1 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 V-ATPase A1 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 A1 Antibody (E-8): sc-374475
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
V-ATPase A1 CRISPR/Cas9 KO Plasmid (h) | sc-400975 | 20 µg | $397.00 |
Overview
ATP6V0A1 encodes the a1 isoform of the V0 “a” subunit within vacuolar H\+-ATPase (V-ATPase), a multi-subunit proton pump that acidifies endosomes, lysosomes, and secretory vesicles. By establishing luminal pH gradients, V-ATPase A1 supports receptor-mediated endocytosis, lysosomal proteolysis, autophagy-lysosome flux, and vesicular trafficking, and it contributes to ion homeostasis and organelle maturation. Perturbation of V-ATPase–dependent acidification is linked to defects in protein turnover and membrane transport, with downstream impacts on cellular stress responses and neurodegenerative and cancer-associated phenotypes. As a core regulator of organelle acidification, V-ATPase A1 is frequently studied in pathways governing nutrient sensing, mTORC1 signaling at lysosomes, and endolysosomal dynamics.
V-ATPase A1 CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ATP6V0A1 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ATP6V0A1 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 ATP6V0A1 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 A1 protein expression.
This CRISPR knockout system enables efficient generation of ATP6V0A1-deficient cell models for investigation of V-ATPase A1 signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting ATP6V0A1 exon(s) critical for V-ATPase A1 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 ATP6V0A1 genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by V-ATPase A1 CRISPR/Cas9 KO Plasmid (h) and V-ATPase A1 CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ATP6V0A1 locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by V-ATPase A1 HDR Plasmid (h) and V-ATPase A1 HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ATP6V0A1 homology arms to support homology-directed repair at defined ATP6V0A1 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.