c-Jun CRISPR Activation Plasmid (m): sc-421207-ACT

Jun encodes the transcription factor c-Jun, a core component of the AP-1 complex that integrates MAPK signaling inputs to control stimulus-responsive gene expression. c-Jun regulates programs involved in cell-cycle progression, apoptosis, differentiation, and inflammatory responses through cooperative binding with FOS family proteins at AP-1 sites. In mouse systems, JUN/AP-1 activity is widely used as a readout of stress and cytokine signaling, linking upstream pathways such as JNK, ERK, and p38 to downstream transcriptional remodeling. Dysregulated c-Jun signaling is implicated in oncogenic transformation, tissue remodeling, and immune-mediated pathology, making Jun a common node for mechanistic studies of signaling-to-transcription coupling.

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

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

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