INCENP CRISPR Activation Plasmid (h): sc-403186-ACT

INCENP (inner centromere protein) is a core component of the chromosomal passenger complex that coordinates mitotic chromosome alignment, spindle assembly checkpoint signaling, and cytokinesis. Through functional coupling with Aurora B kinase, Survivin (BIRC5), and Borealin (CDCA8), INCENP helps regulate kinetochore–microtubule attachments, histone H3 phosphorylation, and correction of erroneous spindle interactions. Proper INCENP activity is therefore essential for faithful chromosome segregation and maintenance of genomic stability during cell division. Dysregulation of chromosomal passenger complex signaling is associated with aneuploidy and proliferative phenotypes relevant to cancer biology and other chromosome instability contexts.

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

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

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