Aquaporin 8/AQP8 CRISPR Activation Plasmid (h): sc-402490-ACT

AQP8 encodes aquaporin 8, a membrane channel that facilitates rapid water transport and also permits passage of small solutes such as hydrogen peroxide, linking membrane permeability to osmotic balance and redox signaling. AQP8 is expressed in epithelial and immune-relevant contexts where it supports fluid movement, organelle homeostasis, and regulated reactive oxygen species flux that can influence pathways controlling proliferation, differentiation, and stress responses. Altered AQP8 expression has been reported across inflammatory states and multiple cancer types, where dysregulated water and H2O2 transport can reshape cell migration, barrier function, and signaling dynamics. As a result, AQP8 is commonly studied in models of epithelial physiology, oxidative stress biology, and tumor cell behavior.

Aquaporin 8/AQP8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AQP8 expression without altering the underlying DNA sequence.

Aquaporin 8/AQP8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AQP8 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 AQP8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Aquaporin 8/AQP8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AQP8 locus and enabling the study of Aquaporin 8/AQP8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Aquaporin 8/AQP8 pathway restoration in tumor cells with silenced or reduced AQP8 expression.

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