MKP-3 CRISPR Activation Plasmid (h): sc-400850-ACT

Dual specificity phosphatase 6 (DUSP6), also known as MKP-3, is a cytosolic MAP kinase phosphatase that preferentially dephosphorylates and inactivates ERK1/2, providing negative feedback control of the RAS–RAF–MEK–ERK signaling cascade. By constraining ERK-driven transcriptional programs, MKP-3 influences proliferation, differentiation, and stimulus-dependent signaling dynamics downstream of receptor tyrosine kinases such as EGFR and FGFR. Altered DUSP6 expression or activity can reshape MAPK pathway output and has been reported in contexts where ERK signaling is dysregulated, including multiple cancer-associated models and developmental signaling studies. As a pathway modulator, DUSP6 is frequently used to interrogate MAPK feedback regulation and signaling adaptation.

MKP-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DUSP6 expression without altering the underlying DNA sequence.

MKP-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DUSP6 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 DUSP6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MKP-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DUSP6 locus and enabling the study of MKP-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MKP-3 pathway restoration in tumor cells with silenced or reduced DUSP6 expression.

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