ERK 3 CRISPR Activation Plasmid (h): sc-403457-ACT

MAPK6 encodes extracellular signal-regulated kinase 3 (ERK3), an atypical MAP kinase that integrates mitogenic and stress-related cues to regulate cell growth, differentiation, migration, and cytoskeletal dynamics. ERK3 signaling is linked to downstream regulation of transcriptional programs and kinase networks that influence cell-cycle progression and motility, including reported functional coupling with kinases such as MK5 in specific contexts. Dysregulated MAPK6/ERK3 expression or activity has been associated with altered proliferative and invasive phenotypes in multiple model systems, supporting its relevance to cancer biology and other diseases involving aberrant signaling and tissue remodeling. As a non-canonical MAPK, ERK3 provides a complementary entry point to study MAPK pathway diversity beyond the classical ERK1/2 cascade.

ERK 3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAPK6 expression without altering the underlying DNA sequence.

ERK 3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAPK6 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 MAPK6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ERK 3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAPK6 locus and enabling the study of ERK 3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ERK 3 pathway restoration in tumor cells with silenced or reduced MAPK6 expression.

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