Fibrocystin CRISPR Activation Plasmid (h): sc-403635-ACT

PKHD1 encodes fibrocystin/polyductin, a large membrane-associated protein enriched at primary cilia and basal bodies in renal and biliary epithelial cells, where it contributes to ciliogenesis, mechanosensory signaling, and maintenance of epithelial polarity. Fibrocystin participates in cilia-linked pathways that coordinate calcium-dependent signaling, planar cell polarity, and tubulogenesis during ductal and nephron development. Disruption or altered expression of PKHD1 is strongly associated with hepatorenal fibrocystic phenotypes, including autosomal recessive polycystic kidney disease, reflecting defects in ciliary signaling and ductal morphogenesis. Experimental modulation of fibrocystin levels is therefore relevant for studying cilia-dependent epithelial differentiation, lumen formation, and stress responses in kidney and liver model systems.

Fibrocystin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PKHD1 expression without altering the underlying DNA sequence.

Fibrocystin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PKHD1 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 PKHD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Fibrocystin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PKHD1 locus and enabling the study of Fibrocystin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Fibrocystin pathway restoration in tumor cells with silenced or reduced PKHD1 expression.

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