Plk CRISPR Activation Plasmid (h): sc-400411-ACT

Human PLK1 encodes polo-like kinase 1 (Plk), a serine/threonine kinase that coordinates mitotic entry and progression by regulating centrosome maturation, bipolar spindle assembly, kinetochore–microtubule attachment, and cytokinesis. PLK1 activity integrates with CDK1–cyclin B signaling and phosphorylation networks controlling chromosome segregation and the spindle assembly checkpoint. Dysregulated PLK1 expression or activity perturbs genomic stability and cell cycle control, features frequently studied in rapidly proliferating cellular states and tumor-associated phenotypes. As a central node in mitotic signaling, PLK1 is widely used as a functional readout in studies of proliferation, checkpoint regulation, and mitosis-linked stress responses.

Plk CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLK1 expression without altering the underlying DNA sequence.

Plk CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLK1 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 PLK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Plk expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLK1 locus and enabling the study of Plk-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Plk pathway restoration in tumor cells with silenced or reduced PLK1 expression.

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