CA XII CRISPR Activation Plasmid (h): sc-404149-ACT

Human CA12 encodes carbonic anhydrase XII (CA XII), a membrane-associated zinc metalloenzyme that catalyzes reversible CO₂ hydration to bicarbonate and protons, supporting extracellular and pericellular pH regulation. By controlling bicarbonate availability and proton flux, CA XII influences ion transport, cell adhesion dynamics, and metabolic adaptation in hypoxic or acidic microenvironments. CA XII activity intersects with pH-dependent signaling and transporter networks, including bicarbonate transporters and proton-coupled processes that shape redox balance and glycolytic physiology. Altered CA12 expression has been reported in multiple disease-relevant contexts where dysregulated pH homeostasis and microenvironmental acidification contribute to cellular stress responses and phenotypic plasticity.

CA XII CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CA12 expression without altering the underlying DNA sequence.

CA XII CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CA12 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the CA12 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CA XII expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CA12 locus and enabling the study of CA XII-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CA XII pathway restoration in tumor cells with silenced or reduced CA12 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.