CA II CRISPR/Cas9 KO Plasmid (h): sc-401059

Overview

Carbonic anhydrase II (CA II), encoded by the human CA2 gene, is a highly active cytosolic metalloenzyme that catalyzes the reversible hydration of CO₂ to bicarbonate and protons, thereby regulating intracellular pH, CO₂ transport, and bicarbonate availability. Through coupling to bicarbonate transporters and proton-handling mechanisms, CA II supports acid–base homeostasis across diverse tissues and contributes to processes such as epithelial ion transport, osteoclast-mediated bone resorption, and erythrocyte gas exchange. CA2 activity intersects with pH-sensitive metabolic regulation and cellular stress responses, where altered carbonic anhydrase function can influence signaling and enzyme activity downstream of proton dynamics. Genetic and functional perturbation of CA2 has been associated with disorders of acid–base balance and mineralization, making it a useful target for mechanistic studies of pH-dependent physiology and disease-relevant cellular phenotypes.

CA II CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the CA2 gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the CA2 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 CA2 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 II protein expression.

This CRISPR knockout system enables efficient generation of CA2-deficient cell models for investigation of CA II signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.

Key Features

• sgRNAs targeting CA2 exon(s) critical for CA II 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 CA2 genomic sites to improve knockout efficiency
• Compatible with delivery by transfection

Design Variants

CRISPRs +/- HDRs

• gRNAs encoded by CA II CRISPR/Cas9 KO Plasmid (h) and CA II CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the CA2 locus. One or both targeting designs may be available. See Related Products for availability.
• HDR donor constructs encoded by CA II HDR Plasmid (h) and CA II HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by CA2 homology arms to support homology-directed repair at defined CA2 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.