CD47 CRISPR Activation Plasmid (h): sc-400508-ACT

Human CD47 encodes a widely expressed cell-surface immunoglobulin superfamily protein that functions as a “self” recognition signal and adhesion receptor. CD47 engages SIRPα on myeloid cells to modulate phagocytosis and immune surveillance, and it also binds thrombospondin-1 to influence integrin signaling, cytoskeletal dynamics, and cell migration. Through these interactions, CD47 contributes to processes including leukocyte trafficking, vascular and platelet biology, and regulation of inflammatory responses. Dysregulated CD47 expression has been associated with altered tumor–immune interactions, hematologic malignancies, and inflammatory or fibrotic pathologies, making it a frequent target for mechanistic studies of immune evasion and tissue microenvironment remodeling.

CD47 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD47 expression without altering the underlying DNA sequence.

CD47 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD47 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 CD47 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD47 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD47 locus and enabling the study of CD47-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD47 pathway restoration in tumor cells with silenced or reduced CD47 expression.

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