PBK CRISPR Activation Plasmid (h): sc-404069-ACT

PDZ binding kinase (PBK/TOPK) is a serine/threonine kinase enriched in proliferative tissues and frequently elevated in transformed cells, where it supports mitotic progression and genome maintenance. PBK functions in cell-cycle control by coordinating G2/M transition and centrosome/spindle dynamics, and it can modulate MAPK signaling to influence stress responses and proliferation programs. Through phosphorylation of substrates involved in chromatin regulation and checkpoint control, PBK contributes to DNA damage tolerance and replicative fitness. Dysregulated PBK expression has been associated with tumor biology, making it a useful node for studying oncogenic signaling, cell-cycle dependency, and proliferation-linked transcriptional states.

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

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

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