CD5 CRISPR Activation Plasmid (h): sc-402309-ACT

CD5 encodes a surface glycoprotein of the scavenger receptor cysteine-rich family expressed primarily on T lymphocytes and a subset of B cells, where it functions as a key modulator of antigen receptor signaling. By associating with the T cell receptor complex and engaging ligands such as CD72, CD5 helps tune activation thresholds, calcium flux, cytokine production, and downstream MAPK and NF-κB signaling outputs that shape immune homeostasis. Altered CD5 expression or signaling has been linked to dysregulated lymphocyte activation and differentiation, with relevance to autoimmune inflammation and hematologic malignancies characterized by aberrant B/T cell phenotypes. As a biomarker and functional regulator, CD5 is frequently used to study immune synapse dynamics, tolerance, and lineage-specific signaling programs.

CD5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD5 expression without altering the underlying DNA sequence.

CD5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD5 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 CD5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD5 locus and enabling the study of CD5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD5 pathway restoration in tumor cells with silenced or reduced CD5 expression.

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