Adipsin CRISPR Activation Plasmid (h): sc-401782-ACT

Complement factor D (CFD), also known as adipsin, is a secreted serine protease that serves as the rate-limiting enzyme of the alternative complement pathway by cleaving factor B bound to C3b to generate the C3 convertase (C3bBb). In addition to innate immune amplification and opsonization, human adipsin is abundantly expressed by adipocytes and links metabolic state to inflammatory signaling within adipose tissue microenvironments. CFD activity influences complement-dependent cytokine programs, immune cell recruitment, and local tissue remodeling through downstream C3a/C5a anaphylatoxin signaling. Dysregulated complement activation and altered CFD expression have been associated with inflammatory and metabolic phenotypes, including adipose dysfunction and broader immune-mediated disease contexts.

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

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

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