Nectin 2 CRISPR Activation Plasmid (h): sc-401994-ACT

NECTIN2 encodes Nectin-2 (CD112), an immunoglobulin-like cell adhesion molecule concentrated at adherens junctions where it mediates Ca2+-independent cell–cell contacts through homophilic and heterophilic interactions. By engaging afadin (AF6) and coordinating with cadherin–catenin complexes, Nectin-2 influences junction assembly, epithelial polarity, and cytoskeletal remodeling during tissue organization and migration. Nectin-2 also participates in immune recognition as a ligand for DNAM-1 (CD226) and TIGIT, linking adhesion biology to NK and T cell signaling at immunological synapses. Dysregulated NECTIN2 expression has been associated with altered adhesion states, immune evasion phenotypes, and progression-related features in multiple malignancies, supporting its investigation in tumor–immune and barrier biology.

Nectin 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NECTIN2 expression without altering the underlying DNA sequence.

Nectin 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NECTIN2 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 NECTIN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nectin 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NECTIN2 locus and enabling the study of Nectin 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nectin 2 pathway restoration in tumor cells with silenced or reduced NECTIN2 expression.

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