C/EBP ζ CRISPR Activation Plasmid (m): sc-419622-ACT

Cebpz encodes the transcription factor C/EBP zeta (C/EBPζ), a member of the CCAAT/enhancer-binding protein family that modulates gene expression programs involved in cell growth control, differentiation, and stress-responsive transcription. C/EBPζ can function as a context-dependent regulator by partnering with other bZIP factors to tune promoter and enhancer activity, linking extracellular cues to changes in RNA polymerase II–driven transcription. In mouse systems, altered C/EBPζ activity is relevant to studies of inflammatory signaling, metabolic adaptation, and cell fate decisions where C/EBP-family networks intersect with cytokine and growth-factor pathways. These properties make Cebpz a useful node for dissecting transcriptional circuitry underlying phenotypic shifts observed in disease-relevant cellular models without implying clinical outcomes.

C/EBP ζ CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cebpz expression without altering the underlying DNA sequence.

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

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