CLCA1 CRISPR Activation Plasmid (h): sc-402998-ACT

Human CLCA1 (chloride channel accessory 1) is a secreted, mucin-associated glycoprotein predominantly expressed in airway and intestinal epithelia, where it modulates epithelial ion transport, mucus hydration, and barrier function. CLCA1 influences calcium-activated chloride conductance indirectly and is linked to goblet cell differentiation and inflammatory signaling programs in mucosal tissues. Its expression tracks with epithelial remodeling processes and has been associated with airway mucus hypersecretion and allergic inflammation, as well as dysregulated epithelial states in gastrointestinal disease contexts. Because CLCA1 is tightly coupled to mucin biology and epithelial differentiation, it serves as a useful marker and mechanistic node for studying secretory lineage transitions and mucosal immune interactions.

CLCA1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CLCA1 expression without altering the underlying DNA sequence.

CLCA1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CLCA1 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 CLCA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CLCA1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CLCA1 locus and enabling the study of CLCA1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CLCA1 pathway restoration in tumor cells with silenced or reduced CLCA1 expression.

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