JNK1 CRISPR Activation Plasmid (h): sc-400048-ACT

MAPK8 encodes the human c-Jun N-terminal kinase 1 (JNK1), a stress-activated MAP kinase that phosphorylates transcription factors such as c-JUN and modulates AP-1–dependent gene programs. JNK1 integrates signals from cytokines, oxidative stress, UV irradiation, and endoplasmic reticulum stress to regulate apoptosis, autophagy, inflammation, and cell cycle control through MAPK and NF-κB crosstalk. This pathway contributes to cellular decisions that influence oncogenic transformation, neurodegenerative processes, and metabolic and inflammatory phenotypes, making MAPK8 a widely used node for mechanistic pathway dissection. Dysregulated JNK1 signaling has been linked to altered immune signaling and stress-response networks in multiple disease-relevant contexts, supporting its utility in functional genomics studies.

JNK1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAPK8 expression without altering the underlying DNA sequence.

JNK1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAPK8 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 MAPK8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous JNK1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAPK8 locus and enabling the study of JNK1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of JNK1 pathway restoration in tumor cells with silenced or reduced MAPK8 expression.

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