PAK6 CRISPR Activation Plasmid (h): sc-403345-ACT

PAK6 (p21-activated kinase 6) is a serine/threonine kinase that functions downstream of Rho family GTPases to coordinate cytoskeletal remodeling, cell motility, and signal-dependent changes in gene expression. It integrates cues from growth factor and stress-responsive pathways, influencing processes such as adhesion dynamics, neurite outgrowth, and survival signaling through kinase-driven phosphorylation networks. In human cells, altered PAK6 activity has been linked to dysregulated proliferative and migratory phenotypes and has been studied in the context of androgen receptor–associated signaling and tumor biology. As a nodal kinase, PAK6 is frequently investigated for its role in pathway rewiring that supports cellular adaptation under oncogenic or microenvironmental stress.

PAK6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PAK6 expression without altering the underlying DNA sequence.

PAK6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PAK6 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 PAK6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PAK6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PAK6 locus and enabling the study of PAK6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PAK6 pathway restoration in tumor cells with silenced or reduced PAK6 expression.

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