MEK Kinase 1 CRISPR Activation Plasmid (h): sc-401107-ACT

Human MAP3K1 encodes MEK Kinase 1 (MEKK1), a serine/threonine MAP kinase kinase kinase that integrates signals from growth factors, cytokines, and cellular stress to regulate downstream MAPK cascades. MEKK1 couples upstream cues to MKK4/7–JNK and MKK3/6–p38 signaling, influencing transcriptional programs that control proliferation, apoptosis, differentiation, and inflammatory responses. Through its regulatory roles in stress-activated kinase pathways and cytoskeletal remodeling, MAP3K1 is frequently studied in contexts of aberrant signaling network rewiring. Altered MAP3K1 activity has been linked to dysregulated MAPK pathway output relevant to oncology, developmental disorders, and immune signaling phenotypes in mechanistic research settings.

MEK Kinase 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAP3K1 expression without altering the underlying DNA sequence.

MEK Kinase 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAP3K1 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 MAP3K1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MEK Kinase 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAP3K1 locus and enabling the study of MEK Kinase 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MEK Kinase 1 pathway restoration in tumor cells with silenced or reduced MAP3K1 expression.

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