MEK-5 CRISPR Activation Plasmid (h): sc-401688-ACT

MAP2K5 encodes mitogen-activated protein kinase kinase 5 (MEK-5), a dual-specificity kinase that functions as the principal upstream activator of ERK5 (MAPK7) in the MEK5–ERK5 signaling axis. This pathway integrates growth factor and stress cues to regulate transcriptional programs controlling proliferation, differentiation, survival, and cytoskeletal dynamics, with documented roles in vascular biology and cellular migration. MEK-5–ERK5 signaling intersects with MAPK-dependent control of immediate-early gene expression and can influence responses to oxidative and inflammatory stimuli. Dysregulation of MAP2K5/ERK5 activity has been associated with altered oncogenic signaling contexts, cardiovascular-related phenotypes, and other conditions where MAPK network rewiring contributes to disease-relevant cellular states.

MEK-5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAP2K5 expression without altering the underlying DNA sequence.

MEK-5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAP2K5 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 MAP2K5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MEK-5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAP2K5 locus and enabling the study of MEK-5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MEK-5 pathway restoration in tumor cells with silenced or reduced MAP2K5 expression.

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