MEK Kinase-4 CRISPR Activation Plasmid (h): sc-404345-ACT

Human MAP3K4 encodes MEK Kinase-4 (also known as MTK1), a MAP kinase kinase kinase that functions upstream of the JNK and p38 MAPK cascades. As a stress- and cytokine-responsive signaling node, MEK Kinase-4 helps couple environmental and developmental cues to transcriptional programs controlling apoptosis, differentiation, and inflammatory responses. MAP3K4 activity contributes to regulation of MAPK-mediated cross-talk with DNA damage signaling and cellular stress adaptation, processes frequently perturbed in cancer biology and immune dysregulation. Dysregulated MAP3K4–JNK/p38 signaling has been implicated in altered cell fate decisions and aberrant responses to oxidative and genotoxic stress, supporting its utility as a pathway modulator in mechanistic studies.

MEK Kinase-4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAP3K4 expression without altering the underlying DNA sequence.

MEK Kinase-4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAP3K4 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 MAP3K4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MEK Kinase-4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAP3K4 locus and enabling the study of MEK Kinase-4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MEK Kinase-4 pathway restoration in tumor cells with silenced or reduced MAP3K4 expression.

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