Pendrin CRISPR Activation Plasmid (h): sc-401272-ACT

Human SLC26A4 encodes pendrin, a multi-pass anion exchanger that transports chloride, iodide, bicarbonate, and thiocyanate across epithelial membranes. Pendrin contributes to ion and pH homeostasis in the inner ear, thyroid, and kidney, linking epithelial transport programs to systemic electrolyte balance. In the thyroid, pendrin supports apical iodide efflux important for iodide handling, while in renal intercalated cells it modulates bicarbonate and chloride exchange involved in acid–base regulation. Dysregulated SLC26A4 activity is associated with inherited hearing impairment and thyroid-related phenotypes, making it a useful target for mechanistic studies of epithelial transport, developmental biology, and genotype–phenotype relationships.

Pendrin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC26A4 expression without altering the underlying DNA sequence.

Pendrin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC26A4 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 SLC26A4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pendrin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC26A4 locus and enabling the study of Pendrin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pendrin pathway restoration in tumor cells with silenced or reduced SLC26A4 expression.

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