KAI 1 CRISPR Activation Plasmid (h): sc-401317-ACT

CD82 (KAI1) is a tetraspanin cell-surface glycoprotein that organizes membrane microdomains and modulates signaling by partnering with integrins, EGFR-family receptors, and other tetraspanins. Through regulation of adhesion, migration, and cytoskeletal dynamics, KAI1 influences processes such as immune cell trafficking, extracellular matrix interactions, and vesicular communication. CD82 has been widely studied in the context of metastasis biology, where altered expression correlates with changes in invasive potential and epithelial–mesenchymal transition-associated phenotypes. Dysregulation of CD82-dependent signaling networks has also been linked to tumor progression and inflammatory microenvironment remodeling in multiple disease models.

KAI 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD82 expression without altering the underlying DNA sequence.

KAI 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD82 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 CD82 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KAI 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD82 locus and enabling the study of KAI 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KAI 1 pathway restoration in tumor cells with silenced or reduced CD82 expression.

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