CLCA2 CRISPR Activation Plasmid (h2): sc-404824-ACT-2

Human CLCA2 (chloride channel accessory 2) encodes a cell-surface/secreted accessory protein implicated in the regulation of chloride conductance and epithelial ion transport, with roles in membrane dynamics and barrier-associated physiology. CLCA2 expression is frequently linked to differentiation and stress-responsive programs, intersecting with signaling processes that influence cell–cell adhesion, migration, and epithelial homeostasis. Dysregulation of CLCA2 has been reported across multiple cancer contexts and other epithelial disorders, supporting its use as a molecular marker for studying tumor-associated phenotypic transitions and microenvironmental responses. Gene editing of CLCA2 enables mechanistic interrogation of chloride-related signaling, epithelial remodeling, and downstream transcriptional networks in human cell models.

CLCA2 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous CLCA2 expression without altering the underlying DNA sequence.

CLCA2 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CLCA2 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 CLCA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CLCA2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CLCA2 locus and enabling the study of CLCA2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CLCA2 pathway restoration in tumor cells with silenced or reduced CLCA2 expression.

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