Nectin 1 CRISPR Activation Plasmid (h): sc-401328-ACT

NECTIN1 encodes nectin-1, an immunoglobulin-like cell adhesion molecule concentrated at adherens junctions that mediates Ca2+-independent homophilic and heterophilic interactions to stabilize cell–cell contacts. Nectin-1 engages afadin (AF6) and coordinates with cadherin–catenin complexes to regulate junction assembly, epithelial polarity, and actin cytoskeleton remodeling, thereby influencing tissue morphogenesis and barrier integrity. Beyond adhesion, nectin-1 contributes to contact-mediated signaling that impacts cell migration and differentiation programs. Altered NECTIN1 function or expression has been linked to developmental abnormalities affecting craniofacial and ectodermal tissues, supporting its relevance in studies of epithelial organization and disease-associated adhesion defects.

Nectin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NECTIN1 expression without altering the underlying DNA sequence.

Nectin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NECTIN1 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 NECTIN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nectin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NECTIN1 locus and enabling the study of Nectin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nectin 1 pathway restoration in tumor cells with silenced or reduced NECTIN1 expression.

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