MEK-2 CRISPR Activation Plasmid (h): sc-402732-ACT

MAP2K2 encodes MEK-2, a dual-specificity MAPK kinase that phosphorylates and activates ERK1/2 to propagate mitogen- and growth factor–responsive signaling. As a core node in the RAS–RAF–MEK–ERK cascade, MEK-2 regulates cell-cycle progression, differentiation, survival, and adaptive transcriptional programs, integrating cues from receptor tyrosine kinases and stress inputs. Dysregulation of MAP2K2/MEK-2 activity contributes to aberrant ERK pathway output implicated in oncogenic signaling, developmental disorders, and altered inflammatory responses. In human model systems, MAP2K2 modulation is commonly used to interrogate pathway dynamics, feedback control, and cross-talk with PI3K/AKT and JNK/p38 signaling.

MEK-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAP2K2 expression without altering the underlying DNA sequence.

MEK-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAP2K2 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 MAP2K2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MEK-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAP2K2 locus and enabling the study of MEK-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MEK-2 pathway restoration in tumor cells with silenced or reduced MAP2K2 expression.

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