CD2 CRISPR Activation Plasmid (h): sc-402105-ACT

Human CD2 encodes a T cell surface adhesion and signaling receptor of the immunoglobulin superfamily that binds CD58 (LFA-3) to stabilize antigen-dependent contact between T lymphocytes and antigen-presenting cells. CD2 participates in immunological synapse formation and modulates TCR signaling thresholds, cytoskeletal reorganization, and downstream pathways that shape activation, cytokine production, and cell–cell communication. Through these roles, CD2 influences lymphocyte development and effector function, and altered CD2 expression or signaling has been associated with dysregulated immune responses in inflammatory and autoimmune settings as well as immune-cell phenotypes studied in hematologic malignancy research. CD2 is therefore a useful node for dissecting mechanisms of T cell activation, adhesion dynamics, and immune microenvironment interactions.

CD2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD2 expression without altering the underlying DNA sequence.

CD2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD2 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 CD2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD2 locus and enabling the study of CD2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD2 pathway restoration in tumor cells with silenced or reduced CD2 expression.

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