γPAK CRISPR Activation Plasmid (h): sc-401595-ACT

PAK2 encodes the human serine/threonine kinase γPAK, a Rac1/Cdc42-responsive effector that coordinates actin cytoskeleton remodeling, focal adhesion turnover, and directional cell motility. γPAK integrates signals from Rho-family GTPases to regulate MAPK-associated transcriptional programs, cell-cycle progression, and stress-response pathways that influence apoptosis and survival. Through phosphorylation of downstream substrates involved in cytoskeletal dynamics and signaling scaffolds, PAK2 helps shape processes such as adhesion-dependent migration and membrane trafficking. Dysregulated PAK2 activity or expression has been linked to altered proliferative and invasive phenotypes in cancer biology and to broader perturbations in signaling networks relevant to inflammatory and neurodevelopmental contexts.

γPAK CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PAK2 expression without altering the underlying DNA sequence.

γPAK CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PAK2 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 PAK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous γPAK expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PAK2 locus and enabling the study of γPAK-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of γPAK pathway restoration in tumor cells with silenced or reduced PAK2 expression.

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