Repo-Man CRISPR Activation Plasmid (h): sc-405105-ACT

CDCA2 encodes Repo-Man, a chromatin-associated targeting subunit that recruits protein phosphatase 1 (PP1) to mitotic chromosomes and the anaphase/telophase nucleus to coordinate dephosphorylation events during mitotic exit. Repo-Man contributes to re-establishment of interphase chromatin architecture and regulation of histone phosphorylation states, linking cell cycle progression with transcriptional and epigenetic control. Through its roles in chromosome segregation, DNA replication timing, and nuclear reassembly, altered CDCA2 activity is frequently studied in the context of proliferative signaling and genome stability. Dysregulation of Repo-Man–PP1–dependent processes is relevant to models of cell cycle checkpoint dysfunction and cancer-associated chromatin remodeling.

Repo-Man CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDCA2 expression without altering the underlying DNA sequence.

Repo-Man CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDCA2 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 CDCA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Repo-Man expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDCA2 locus and enabling the study of Repo-Man-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Repo-Man pathway restoration in tumor cells with silenced or reduced CDCA2 expression.

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