CD46 CRISPR Activation Plasmid (h): sc-400968-ACT

Human CD46 (membrane cofactor protein) is a widely expressed complement regulatory glycoprotein that protects host cells from complement-mediated injury by serving as a cofactor for factor I–dependent cleavage of C3b and C4b. Beyond complement control, CD46 influences cell signaling and immune modulation, including regulation of T cell activation and differentiation and integration with inflammatory pathways. CD46 also functions as a cellular receptor for multiple pathogens, linking its expression to host–pathogen interactions and mucosal immunity. Dysregulated CD46 activity or expression is associated with complement-driven pathology and immune dysfunction, making it a useful target for mechanistic studies in immunology, infection biology, and cell surface signaling.

CD46 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD46 expression without altering the underlying DNA sequence.

CD46 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD46 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 CD46 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD46 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD46 locus and enabling the study of CD46-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD46 pathway restoration in tumor cells with silenced or reduced CD46 expression.

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