Borealin CRISPR Activation Plasmid (h): sc-403003-ACT

CDCA8 encodes Borealin, an essential chromosomal passenger complex (CPC) subunit that coordinates mitotic progression by regulating chromosome alignment, kinetochore–microtubule attachments, spindle checkpoint signaling, and cytokinesis. Through functional coupling with Aurora B kinase and INCENP/Survivin, Borealin helps position CPC activity at centromeres and the midzone to ensure accurate sister chromatid segregation. Perturbation of CDCA8 expression disrupts mitotic fidelity, promotes aneuploidy, and is frequently linked to proliferative states observed across multiple cancer contexts. As a result, CDCA8 is widely studied in pathways governing cell cycle control, chromosomal instability, and stress responses during division.

Borealin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDCA8 expression without altering the underlying DNA sequence.

Borealin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDCA8 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 CDCA8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Borealin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDCA8 locus and enabling the study of Borealin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Borealin pathway restoration in tumor cells with silenced or reduced CDCA8 expression.

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