URB CRISPR Activation Plasmid (h): sc-408597-ACT

CCDC80 encodes URB, an extracellular matrix–associated and secreted protein implicated in stromal remodeling, cell–matrix adhesion, and regulation of growth factor signaling. URB has been linked to pathways that influence adipogenesis, metabolic homeostasis, and context-dependent control of cell proliferation and migration, consistent with a role in tissue organization and mesenchymal–epithelial crosstalk. Altered CCDC80 expression has been reported across multiple disease contexts, including obesity-related phenotypes and cancers, where it may correlate with changes in tumor microenvironment composition and invasive behavior. These characteristics make CCDC80 a useful target for dissecting extracellular signaling networks, matrix biology, and transcriptional programs associated with phenotypic state changes.

URB CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCDC80 expression without altering the underlying DNA sequence.

URB CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCDC80 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 CCDC80 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous URB expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCDC80 locus and enabling the study of URB-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of URB pathway restoration in tumor cells with silenced or reduced CCDC80 expression.

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