CD6 CRISPR Activation Plasmid (h): sc-403748-ACT

CD6 is a T cell surface glycoprotein of the scavenger receptor cysteine-rich (SRCR) family that modulates antigen receptor signaling at the immunological synapse. Through interactions with ligands such as ALCAM (CD166), CD6 contributes to T cell activation thresholds, adhesion-dependent signaling, and downstream pathways including MAPK and NF-κB that shape cytokine production and effector differentiation. Altered CD6 expression or signaling has been linked to dysregulated immune responses and inflammatory pathology, making it a useful node for studying immune homeostasis and leukocyte communication. In human systems, CD6 provides a tractable handle for dissecting T cell–APC contact dynamics, co-stimulatory integration, and immune network perturbations relevant to autoimmunity and immune-mediated disease models.

CD6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD6 expression without altering the underlying DNA sequence.

CD6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD6 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 CD6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD6 locus and enabling the study of CD6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD6 pathway restoration in tumor cells with silenced or reduced CD6 expression.

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