ERK 2 CRISPR Activation Plasmid (h): sc-400043-ACT

MAPK1 encodes extracellular signal‑regulated kinase 2 (ERK2), a serine/threonine kinase that functions as a central effector of the RAS–RAF–MEK–ERK MAPK cascade. ERK2 integrates growth factor and cytokine inputs to regulate transcriptional programs controlling proliferation, differentiation, survival, and stress responses through phosphorylation of cytosolic and nuclear substrates. This pathway also coordinates cell-cycle progression and feedback regulation of upstream signaling, shaping signal amplitude and duration. Dysregulated ERK signaling is widely implicated in oncogenic transformation and altered developmental and inflammatory processes, making MAPK1/ERK2 a key node for mechanistic studies of signaling dynamics and pathway cross-talk.

ERK 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAPK1 expression without altering the underlying DNA sequence.

ERK 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAPK1 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 MAPK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ERK 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAPK1 locus and enabling the study of ERK 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ERK 2 pathway restoration in tumor cells with silenced or reduced MAPK1 expression.

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