ATP6F CRISPR/Cas9 KO Plasmid (h): sc-406611
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ATP6F 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 ATP6F 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
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ATP6F CRISPR/Cas9 KO Plasmid (h) | sc-406611 | 20 µg | $397.00 |
Overview
ATP6V0B encodes ATP6F, an accessory subunit of the vacuolar H+-ATPase (V-ATPase) that supports ATP-driven proton translocation to acidify endosomes, lysosomes, and secretory vesicles. V-ATPase activity is central to endocytic trafficking, receptor recycling, autophagy-lysosome function, and pH-dependent processing of macromolecules, thereby influencing nutrient sensing and cellular stress responses. Perturbation of V-ATPase subunits can disrupt organelle homeostasis and has been associated with altered vesicle dynamics and disease-relevant phenotypes in contexts such as neurodegeneration, infection biology, and cancer cell metabolism, where compartmental acidification is frequently remodeled.
ATP6F CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the ATP6V0B gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the ATP6V0B 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 ATP6V0B 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 ATP6F protein expression.
This CRISPR knockout system enables efficient generation of ATP6V0B-deficient cell models for investigation of ATP6F signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting ATP6V0B exon(s) critical for ATP6F 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 ATP6V0B genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by ATP6F CRISPR/Cas9 KO Plasmid (h) and ATP6F CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the ATP6V0B locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by ATP6F HDR Plasmid (h) and ATP6F HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by ATP6V0B homology arms to support homology-directed repair at defined ATP6V0B 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.