FHR-1 CRISPR Activation Plasmid (h): sc-416510-ACT

CFHR1 encodes complement factor H–related protein 1 (FHR-1), a soluble regulator of the alternative pathway of complement that modulates C3 convertase activity and shapes amplification of complement activation on host and microbial surfaces. By competing with complement factor H and engaging complement components, FHR-1 influences opsonization, inflammatory signaling, and immune complex handling, linking innate immune surveillance with tissue homeostasis. Genetic variation or altered expression of CFHR1 is associated with complement dysregulation phenotypes and has been studied in contexts such as atypical hemolytic uremic syndrome, C3 glomerulopathy, age-related macular degeneration, and other inflammation-driven renal and ocular pathologies. As a result, CFHR1 is widely used in mechanistic studies of complement control, extracellular immune regulation, and downstream injury pathways.

FHR-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CFHR1 expression without altering the underlying DNA sequence.

FHR-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CFHR1 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 CFHR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FHR-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CFHR1 locus and enabling the study of FHR-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FHR-1 pathway restoration in tumor cells with silenced or reduced CFHR1 expression.

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