KIF20A CRISPR Activation Plasmid (h): sc-403194-ACT

KIF20A (also known as MKLP2) encodes a kinesin-like motor protein that associates with microtubules to coordinate mitotic progression, particularly during anaphase and cytokinesis. It supports central spindle organization and abscission through regulation of chromosomal passenger complex localization and key mitotic kinases, linking KIF20A activity to cell-cycle control and genome stability pathways. Dysregulated KIF20A expression has been reported across multiple tumor contexts and is frequently used as a proliferation-associated marker in studies of oncogenic cell division programs. As a result, KIF20A is broadly relevant for investigating mitotic fidelity, spindle dynamics, and mechanisms that couple cytokinesis to aneuploidy and cellular stress responses.

KIF20A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KIF20A expression without altering the underlying DNA sequence.

KIF20A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KIF20A 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 KIF20A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KIF20A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KIF20A locus and enabling the study of KIF20A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KIF20A pathway restoration in tumor cells with silenced or reduced KIF20A expression.

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