ERK 1 CRISPR Activation Plasmid (h): sc-400010-ACT

MAPK3 encodes extracellular signal–regulated kinase 1 (ERK1), a serine/threonine kinase that functions as a core effector of the RAS–RAF–MEK–ERK MAPK cascade. ERK1 transduces signals from growth factors, cytokines, and stress stimuli to regulate transcriptional programs controlling proliferation, differentiation, metabolism, and cell survival through phosphorylation of nuclear and cytoplasmic substrates. This pathway coordinates cell-cycle progression, immediate early gene expression, and feedback regulation of upstream receptors and kinases. Dysregulated ERK signaling is frequently implicated in oncogenic signaling networks and inflammatory or neurobiological phenotypes, making MAPK3 a common node for mechanistic studies of pathway rewiring and context-dependent signaling outputs.

ERK 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAPK3 expression without altering the underlying DNA sequence.

ERK 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAPK3 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 MAPK3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ERK 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAPK3 locus and enabling the study of ERK 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ERK 1 pathway restoration in tumor cells with silenced or reduced MAPK3 expression.

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