Cdc27 CRISPR Activation Plasmid (h): sc-400789-ACT

CDC27 encodes Cdc27, a core subunit of the anaphase-promoting complex/cyclosome (APC/C), an E3 ubiquitin ligase that drives ordered proteolysis of key mitotic regulators to ensure faithful chromosome segregation and cell-cycle transitions. Through APC/C-dependent ubiquitination of substrates such as securin and cyclins, Cdc27 helps coordinate spindle checkpoint signaling, mitotic exit, and maintenance of genomic stability. Dysregulation of APC/C components and checkpoint control is associated with chromosomal instability and proliferative phenotypes observed across multiple tumor contexts, making CDC27 a useful node for studying mitotic control and ubiquitin–proteasome system dynamics. Human CDC27 is therefore widely applied in investigations of cell division timing, checkpoint fidelity, and pathways linking proteostasis to genome maintenance.

Cdc27 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDC27 expression without altering the underlying DNA sequence.

Cdc27 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDC27 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 CDC27 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cdc27 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDC27 locus and enabling the study of Cdc27-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cdc27 pathway restoration in tumor cells with silenced or reduced CDC27 expression.

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