C3aR CRISPR Activation Plasmid (h): sc-401448-ACT

C3AR1 encodes the complement component 3a receptor 1 (C3aR), a G protein–coupled receptor that binds the anaphylatoxin C3a to couple complement activation to cellular signaling. C3aR engagement triggers pathways including Gαi/Gαq-mediated calcium flux, MAPK/ERK, and PI3K signaling, shaping leukocyte chemotaxis, cytokine release, and myeloid cell activation. This receptor integrates innate immune sensing with inflammatory amplification and can influence endothelial activation and tissue remodeling. Dysregulated C3AR1 activity has been implicated in inflammatory and immune-mediated disease biology, supporting its study in models of chronic inflammation and microenvironment-driven pathology.

C3aR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous C3AR1 expression without altering the underlying DNA sequence.

C3aR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the C3AR1 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 C3AR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous C3aR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native C3AR1 locus and enabling the study of C3aR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of C3aR pathway restoration in tumor cells with silenced or reduced C3AR1 expression.

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