MEK-6 CRISPR Activation Plasmid (h): sc-402544-ACT

MAP2K6 encodes the human dual-specificity kinase MEK-6 (MKK6), a core component of the MAPK cascade that phosphorylates and activates p38 MAPKs in response to inflammatory cytokines, oxidative stress, and environmental stressors. Through p38 signaling, MEK-6 influences transcriptional programs controlling innate immune responses, apoptosis, differentiation, and cell-cycle checkpoints. Dysregulated MAP2K6–p38 pathway activity has been linked to altered cytokine production and stress-adaptive phenotypes observed across inflammatory disorders and multiple cancer contexts. As an upstream node, MEK-6 provides a tractable handle for dissecting pathway routing and stimulus-specific signal integration.

MEK-6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAP2K6 expression without altering the underlying DNA sequence.

MEK-6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAP2K6 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 MAP2K6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MEK-6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAP2K6 locus and enabling the study of MEK-6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MEK-6 pathway restoration in tumor cells with silenced or reduced MAP2K6 expression.

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