Parafibromin CRISPR Activation Plasmid (h): sc-401334-ACT

CDC73 encodes parafibromin, a nuclear tumor suppressor that functions as a core component of the PAF1 transcriptional regulatory complex, coordinating RNA polymerase II elongation and chromatin-associated gene control. Parafibromin contributes to epigenetic regulation and cell-cycle governance by influencing histone modifications and transcriptional programs linked to proliferation and differentiation. Through these roles, CDC73 helps maintain genomic stability and balanced transcriptional output across multiple signaling contexts, including pathways intersecting with Wnt/β-catenin-responsive gene expression. Disruption of CDC73 function is associated with endocrine and parathyroid-related neoplasia and broader dysregulation of growth control, making it a valuable target for mechanistic studies in cancer biology and transcriptional regulation.

Parafibromin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDC73 expression without altering the underlying DNA sequence.

Parafibromin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDC73 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 CDC73 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Parafibromin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDC73 locus and enabling the study of Parafibromin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Parafibromin pathway restoration in tumor cells with silenced or reduced CDC73 expression.

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