Cochlin CRISPR Activation Plasmid (h): sc-405944-ACT

COCH encodes cochlin, a secreted extracellular matrix glycoprotein enriched in inner ear tissues where it contributes to matrix organization, cell–matrix adhesion, and mechanosensory homeostasis. Cochlin contains LCCL and von Willebrand factor A–like domains that support protein–protein interactions and can influence extracellular proteolysis and inflammatory signaling in local microenvironments. Altered COCH expression or variants are associated with hereditary auditory and vestibular dysfunction, linking cochlin biology to pathways governing extracellular matrix remodeling and stress responses. These features make COCH a useful model for studying how secreted matrix components regulate tissue integrity and sensory organ physiology.

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

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

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