CA V CRISPR/Cas9 KO Plasmid (m): sc-419455
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CA V 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 CA V 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: CA V Antibody (A-1): sc-398184
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CA V CRISPR/Cas9 KO Plasmid (m) | sc-419455 | 20 µg | $397.00 |
Overview
Car5a encodes mitochondrial carbonic anhydrase V (CA V), a zinc metalloenzyme that catalyzes reversible CO₂ hydration to generate bicarbonate within the mitochondrial matrix. This bicarbonate supply supports anaplerotic and biosynthetic reactions by feeding carboxylation-dependent enzymes, linking Car5a to metabolic pathways such as gluconeogenesis, ureagenesis, and lipid and amino acid metabolism. In mouse tissues with high metabolic demand, CA V activity contributes to regulation of mitochondrial pH and carbon flux, thereby influencing cellular energy balance and metabolite homeostasis. Altered mitochondrial carbonic anhydrase function has been associated with metabolic dysregulation and can be studied in the context of disorders affecting nitrogen handling and intermediary metabolism.
CA V CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Car5a gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Car5a 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 Car5a 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 CA V protein expression.
This CRISPR knockout system enables efficient generation of Car5a-deficient cell models for investigation of CA V signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Car5a exon(s) critical for CA V 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 Car5a genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by CA V CRISPR/Cas9 KO Plasmid (m) and CA V CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Car5a locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by CA V HDR Plasmid (m) and CA V HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Car5a homology arms to support homology-directed repair at defined Car5a 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.