C/EBP ε CRISPR Activation Plasmid (h): sc-402310-ACT

CEBPE encodes C/EBP ε, a myeloid-restricted bZIP transcription factor that coordinates late-stage granulopoiesis and neutrophil functional maturation. By binding CCAAT/enhancer elements and cooperating with lineage regulators such as GFI1 and SPI1/PU.1, C/EBP ε modulates transcriptional programs controlling cell-cycle exit, granule protein expression, and innate immune effector pathways. Perturbation of CEBPE expression or activity disrupts neutrophil differentiation and is linked to defective granulocyte function, making it a useful node for studying hematopoietic development and inflammatory signaling. In cancer and immune biology models, altered CEBPE-regulated transcription can serve as a readout for dysregulated myeloid lineage commitment and maturation state.

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

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

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