C/EBP beta CRISPR Activation Plasmid (m): sc-419623-ACT

Mouse Cebpb encodes C/EBP beta, a basic leucine zipper transcription factor that coordinates gene programs controlling myeloid differentiation, adipogenesis, and hepatocyte acute-phase responses. It integrates signaling downstream of inflammatory cytokines and growth factors to regulate cell-cycle progression, survival, and metabolic remodeling, with prominent roles in macrophage polarization and stress-responsive transcription. C/EBP beta activity intersects with pathways such as NF-κB, MAPK/ERK, JAK/STAT, and TGF-β, shaping chromatin accessibility and enhancer usage in lineage-committed cells. Dysregulated Cebpb expression or activity has been linked to inflammatory phenotypes, metabolic dysfunction, and tumor-associated transcriptional states, making it a useful node for mechanistic studies of immune–metabolic crosstalk and transcriptional control.

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

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

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