Prestin CRISPR Activation Plasmid (h): sc-401428-ACT

SLC26A5 encodes prestin, a membrane-bound anion transporter–like motor protein that is highly enriched in cochlear outer hair cells and converts changes in membrane potential into rapid electromotility. This electromechanical transduction supports cochlear amplification and sharp frequency tuning, integrating ion homeostasis with voltage-dependent conformational cycling at the plasma membrane. Prestin activity is linked to epithelial ion transport processes and the specialized cytoskeletal and membrane mechanics required for auditory function. Altered SLC26A5 expression or function is associated with sensorineural hearing phenotypes and is used as a molecular entry point to study hair-cell biomechanics and auditory pathway dysregulation.

Prestin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC26A5 expression without altering the underlying DNA sequence.

Prestin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC26A5 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 SLC26A5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Prestin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC26A5 locus and enabling the study of Prestin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Prestin pathway restoration in tumor cells with silenced or reduced SLC26A5 expression.

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