MKP-7 CRISPR Activation Plasmid (h): sc-405727-ACT

DUSP16 encodes mitogen-activated protein kinase phosphatase 7 (MKP-7), a dual-specificity phosphatase that attenuates MAPK signaling by dephosphorylating stress-activated kinases, with reported preference toward JNK and p38 pathways. Through negative feedback control of MAPK cascades, MKP-7 influences stimulus-dependent transcriptional programs that govern proliferation, differentiation, and apoptosis. Dysregulated DUSP16 activity has been linked in the literature to altered inflammatory signaling and tumor-associated MAPK network remodeling, making it a useful node for studying pathway homeostasis. As a cytosolic regulator of kinase-driven responses, MKP-7 is frequently examined in contexts of cytokine signaling, cellular stress adaptation, and immune-related pathway crosstalk.

MKP-7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DUSP16 expression without altering the underlying DNA sequence.

MKP-7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DUSP16 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 DUSP16 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MKP-7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DUSP16 locus and enabling the study of MKP-7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MKP-7 pathway restoration in tumor cells with silenced or reduced DUSP16 expression.

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