CA XIV CRISPR Activation Plasmid (h): sc-404085-ACT

Human CA14 encodes carbonic anhydrase XIV (CA XIV), a membrane-associated zinc metalloenzyme that catalyzes reversible hydration of CO₂ to bicarbonate and protons, thereby regulating extracellular and pericellular pH. By shaping bicarbonate flux and proton availability, CA XIV functionally couples to ion transport systems such as bicarbonate transporters and Na⁺/H⁺ exchangers, influencing epithelial and neuronal acid–base homeostasis. Altered carbonic anhydrase activity and pH dysregulation are linked to processes including cell migration, synaptic physiology, and metabolic adaptation, making CA14 a relevant target for studying microenvironmental control in disease-associated contexts.

CA XIV CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CA14 expression without altering the underlying DNA sequence.

CA XIV CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CA14 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 CA14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CA XIV expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CA14 locus and enabling the study of CA XIV-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CA XIV pathway restoration in tumor cells with silenced or reduced CA14 expression.

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