Cdc34 CRISPR Activation Plasmid (h): sc-403783-ACT

CDC34 encodes the human E2 ubiquitin-conjugating enzyme Cdc34, a core component of SCF (SKP1–CUL1–F-box) E3 ubiquitin ligase complexes that catalyze polyubiquitination of regulatory proteins for proteasomal degradation. Cdc34 activity is tightly linked to cell cycle progression, particularly G1/S control, by promoting turnover of key substrates that regulate DNA replication and checkpoint signaling. Through its role in ubiquitin-dependent proteostasis, CDC34 connects to pathways governing proliferation, genome stability, and stress responses. Altered CDC34 expression or SCF axis dysregulation is frequently studied in the context of oncogenic signaling and other diseases characterized by aberrant protein degradation programs.

Cdc34 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDC34 expression without altering the underlying DNA sequence.

Cdc34 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDC34 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 CDC34 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cdc34 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDC34 locus and enabling the study of Cdc34-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cdc34 pathway restoration in tumor cells with silenced or reduced CDC34 expression.

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