C9 CRISPR Activation Plasmid (h): sc-401198-ACT

Human C9 encodes complement component 9, a terminal effector of the complement cascade that polymerizes within the membrane attack complex (MAC) to perforate target membranes and promote lytic injury. C9 function integrates with innate immune signaling through classical, lectin, and alternative pathway activation, shaping inflammation, opsonization, and pathogen clearance. Dysregulated complement terminal pathway activity and MAC deposition have been implicated in tissue damage and chronic inflammation across immune-mediated and degenerative conditions, supporting the use of C9 as a mechanistic node for studying complement-driven pathology. Experimental modulation of C9 expression helps interrogate how terminal complement activity influences cellular stress responses, barrier integrity, and immune cell–tissue interactions.

C9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous C9 expression without altering the underlying DNA sequence.

C9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the C9 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 C9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous C9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native C9 locus and enabling the study of C9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of C9 pathway restoration in tumor cells with silenced or reduced C9 expression.

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