CD3-γ CRISPR Activation Plasmid (h): sc-402537-ACT

CD3G encodes CD3-γ, an essential component of the T cell receptor (TCR)-CD3 complex that couples antigen recognition to intracellular signal transduction. Through assembly with CD3δ, CD3ε, and CD3ζ, CD3-γ helps initiate phosphorylation-dependent signaling cascades involving LCK, ZAP70, and downstream MAPK, NF-κB, and NFAT pathways that regulate T cell activation, proliferation, and cytokine production. Altered CD3G expression or function can perturb thymocyte development and TCR signaling thresholds, linking this gene to immune dysregulation phenotypes and immunodeficiency-related mechanisms. As a core TCR complex subunit, CD3-γ is widely studied in lymphocyte biology, adaptive immune signaling, and pathway cross-talk that shapes responses to inflammatory cues.

CD3-γ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD3G expression without altering the underlying DNA sequence.

CD3-γ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD3G 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 CD3G transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD3-γ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD3G locus and enabling the study of CD3-γ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD3-γ pathway restoration in tumor cells with silenced or reduced CD3G expression.

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