CD69 CRISPR Activation Plasmid (m2): sc-419566-ACT-2

Mouse Cd69 encodes CD69, an inducible C-type lectin–like surface receptor rapidly upregulated following antigen receptor or cytokine stimulation, serving as an early marker of lymphocyte activation. CD69 modulates immune cell trafficking and tissue retention, in part through regulation of sphingosine-1-phosphate receptor signaling, and influences downstream activation programs including NF-κB- and MAPK-associated responses that shape T cell and NK cell effector function. Altered CD69 activity has been linked to immune dysregulation in models of inflammation, autoimmunity, and tumor immunology, reflecting its role in balancing activation and migration cues. Cd69 gene editing in mouse systems supports mechanistic studies of activation kinetics, tissue-resident immune cell biology, and pathway perturbation analyses in primary cells or in vivo immunology models.

CD69 CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Cd69 expression without altering the underlying DNA sequence.

CD69 CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Cd69 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 Cd69 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD69 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Cd69 locus and enabling the study of CD69-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD69 pathway restoration in tumor cells with silenced or reduced Cd69 expression.

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