cyclin C CRISPR Activation Plasmid (h): sc-402308-ACT

Human CCNC encodes cyclin C, a transcriptional cyclin that partners with CDK8 or CDK19 within the Mediator kinase module to tune RNA polymerase II–dependent transcription. Through regulation of stimulus-responsive gene programs, cyclin C influences cell-cycle progression, metabolic adaptation, and stress signaling, including pathways that govern mitochondrial dynamics and apoptosis. CCNC activity intersects with WNT/β-catenin, TGF-β, and NF-κB transcriptional networks, linking Mediator kinase signaling to developmental and inflammatory processes. Dysregulated CCNC/CDK8–Mediator function has been associated with altered transcriptional states observed in cancer and other diseases characterized by perturbed gene expression control.

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

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

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