Sororin CRISPR Activation Plasmid (h): sc-403556-ACT

CDCA5 encodes Sororin, a conserved regulator of sister chromatid cohesion that stabilizes cohesin complexes on replicated DNA during S phase and ensures accurate chromosome segregation in mitosis. Sororin activity is coordinated with cell-cycle signaling, including APC/C-driven proteolysis and antagonism of WAPL-mediated cohesin release, linking it to replication-coupled cohesion establishment and maintenance. Through its central role in genome stability, Sororin supports faithful DNA repair and replication stress responses, processes frequently perturbed in proliferative disorders. Altered CDCA5 expression has been associated with dysregulated cell-cycle progression and aneuploidy-related phenotypes, making it a useful target for mechanistic studies of chromosomal instability.

Sororin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDCA5 expression without altering the underlying DNA sequence.

Sororin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDCA5 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 CDCA5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sororin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDCA5 locus and enabling the study of Sororin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sororin pathway restoration in tumor cells with silenced or reduced CDCA5 expression.

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