CD5 CRISPR Activation Plasmid (m): sc-419560-ACT

Mouse Cd5 encodes CD5, a surface glycoprotein of the scavenger receptor cysteine-rich family that modulates antigen receptor signaling in T cells and a subset of B cells. CD5 participates in immunological synapse organization and calibrates activation thresholds by integrating signals downstream of the T cell receptor and associated Src-family kinase pathways, influencing cytokine production and lymphocyte survival. Through its role in maintaining immune homeostasis and peripheral tolerance, altered CD5 expression or signaling has been associated with dysregulated lymphocyte activation and inflammatory immune phenotypes. Cd5 is therefore a useful molecular handle for studying signaling feedback, lymphocyte differentiation, and mechanisms that shape adaptive immune responses in mouse models.

CD5 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cd5 expression without altering the underlying DNA sequence.

CD5 CRISPR Activation Plasmid (m) 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.