ZPK CRISPR Activation Plasmid (h): sc-404802-ACT

Human MAP3K12 encodes ZPK (also known as DLK), a MAP kinase kinase kinase that functions upstream of the JNK and p38 MAPK cascades to regulate stress-responsive transcriptional programs. ZPK integrates signals from axonal injury, inflammatory cues, and cytoskeletal perturbations to influence neuronal differentiation, survival, and degeneration, with prominent roles in the nervous system. Through phosphorylation of downstream MAP2Ks, MAP3K12 helps control apoptosis, neurite outgrowth, and synaptic remodeling. Dysregulated ZPK signaling has been associated with neurodegeneration and neuropathic pain biology, making MAP3K12 a useful node for dissecting kinase-driven stress signaling in human cell models.

ZPK CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAP3K12 expression without altering the underlying DNA sequence.

ZPK CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAP3K12 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 MAP3K12 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZPK expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAP3K12 locus and enabling the study of ZPK-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZPK pathway restoration in tumor cells with silenced or reduced MAP3K12 expression.

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