CD40 CRISPR Activation Plasmid (m): sc-423444-ACT

Mouse Cd40 encodes CD40, a TNF receptor superfamily costimulatory molecule expressed on antigen-presenting cells that coordinates adaptive immunity through CD40–CD40L interactions. CD40 signaling activates NF-κB, MAPK, and PI3K pathways to drive dendritic cell maturation, B cell proliferation, immunoglobulin class-switch recombination, and germinal center formation. Through crosstalk with cytokine and pattern-recognition receptor programs, CD40 shapes T cell priming and inflammatory polarization. Dysregulated CD40 activity is implicated in models of autoimmunity, chronic inflammation, transplant rejection biology, and tumor–immune microenvironment regulation, making it a key node for mechanistic immunology studies in mouse systems.

CD40 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cd40 expression without altering the underlying DNA sequence.

CD40 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Cd40 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 Cd40 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD40 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Cd40 locus and enabling the study of CD40-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD40 pathway restoration in tumor cells with silenced or reduced Cd40 expression.

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