CGNL1 CRISPR Activation Plasmid (h): sc-405346-ACT

CGNL1 (cingulin-like 1) encodes a tight junction–associated scaffolding protein that localizes to epithelial and endothelial cell–cell contacts and helps organize junctional architecture. It links junctional complexes to the actin cytoskeleton and participates in regulation of barrier function, polarity, and junction assembly through pathways involving Rho-family GTPase signaling and cytoskeletal remodeling. By influencing paracellular permeability and contact-dependent signaling, CGNL1 is relevant to studies of tissue integrity, inflammation-associated barrier disruption, and microenvironmental cues that shape cell migration. Altered junctional regulation and cytoskeletal dynamics connected to CGNL1 biology are frequently examined in contexts such as epithelial-to-mesenchymal transitions, vascular permeability, and cancer-associated changes in adhesion.

CGNL1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CGNL1 expression without altering the underlying DNA sequence.

CGNL1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CGNL1 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 CGNL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CGNL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CGNL1 locus and enabling the study of CGNL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CGNL1 pathway restoration in tumor cells with silenced or reduced CGNL1 expression.

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