PRAK CRISPR Activation Plasmid (h): sc-403215-ACT

Human MAPKAPK5 encodes PRAK (p38-regulated/activated kinase), a serine/threonine kinase activated downstream of stress-responsive MAPK signaling, particularly the p38 MAPK axis. PRAK participates in phosphorylation programs that shape cellular responses to inflammatory cues, oxidative stress, and DNA damage, with reported roles in regulating transcriptional networks, cytoskeletal dynamics, and cell cycle checkpoints. Through these functions, MAPKAPK5/PRAK is frequently studied in contexts where stress signaling rewires growth and survival programs, including cancer biology and chronic inflammation–associated phenotypes. Its pathway connectivity makes it a useful node for dissecting MAPK-driven signaling crosstalk and adaptive stress responses in human cells.

PRAK CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAPKAPK5 expression without altering the underlying DNA sequence.

PRAK CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAPKAPK5 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 MAPKAPK5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRAK expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAPKAPK5 locus and enabling the study of PRAK-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRAK pathway restoration in tumor cells with silenced or reduced MAPKAPK5 expression.

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