PRK2 CRISPR Activation Plasmid (h): sc-407649-ACT

PKN2 encodes protein kinase N2 (PRK2), a serine/threonine kinase of the AGC family that functions as an effector of Rho family GTPase signaling to coordinate cytoskeletal dynamics, focal adhesion turnover, and cell motility. PRK2 activity integrates cues from GPCR- and growth factor–linked pathways to influence actin remodeling, stress fiber formation, and vesicle trafficking, with downstream effects on proliferation and survival programs. Through these roles, PRK2 has been implicated in processes relevant to tumor cell invasion and metastatic behavior, as well as broader dysregulation of migration and adhesion-dependent signaling observed in diverse disease contexts. Modulating endogenous PKN2 expression is therefore useful for dissecting pathway connectivity between Rho/ROCK-associated signaling, cytoskeletal organization, and transcriptional outputs.

PRK2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PKN2 expression without altering the underlying DNA sequence.

PRK2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PKN2 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 PKN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRK2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PKN2 locus and enabling the study of PRK2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRK2 pathway restoration in tumor cells with silenced or reduced PKN2 expression.

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