JunB CRISPR Activation Plasmid (h): sc-400493-ACT

JUNB encodes JunB, an AP-1 family transcription factor that rapidly responds to mitogenic and stress cues to regulate gene expression programs controlling proliferation, differentiation, and apoptosis. JunB integrates signaling from MAPK/ERK and JNK pathways and cooperates with Fos proteins to modulate enhancer and promoter activity across immune and stromal cell contexts. In human biology, altered JUNB activity has been associated with dysregulated inflammatory signaling, aberrant hematopoietic differentiation, and oncogenic transcriptional networks, making it a useful node for studying context-dependent transcriptional control. JUNB is also frequently used as a readout and regulator in cytokine-driven responses, cell cycle transitions, and chromatin-state remodeling.

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

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

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