ARK-2 CRISPR Activation Plasmid (h): sc-402041-ACT

Human AURKB encodes Aurora kinase B (ARK-2), a serine/threonine kinase that functions as a core catalytic component of the chromosomal passenger complex to coordinate mitotic progression. ARK-2 regulates kinetochore–microtubule attachment, spindle assembly checkpoint signaling, chromosome condensation, and cytokinesis through phosphorylation of substrates such as histone H3 and centromeric proteins. By controlling error correction and chromosome segregation fidelity, AURKB helps maintain genome stability and influences cell-cycle transcriptional programs. Dysregulated AURKB activity is frequently associated with proliferative phenotypes, aneuploidy, and altered mitotic checkpoint control, making it a key node for studying cell division–linked disease mechanisms.

ARK-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AURKB expression without altering the underlying DNA sequence.

ARK-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AURKB 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 AURKB transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ARK-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AURKB locus and enabling the study of ARK-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ARK-2 pathway restoration in tumor cells with silenced or reduced AURKB expression.

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