EBF4 CRISPR Activation Plasmid (h): sc-402438-ACT

EBF4 (early B-cell factor 4) encodes a helix–loop–helix transcription factor within the EBF family that binds DNA and coordinates lineage-restricted gene expression programs. Although less characterized than other EBF paralogs, EBF4 is implicated in regulating transcriptional networks linked to cell differentiation, developmental patterning, and maintenance of cell identity through enhancer and promoter occupancy. By shaping chromatin-accessible states and transcriptional outputs, EBF4 can influence processes such as proliferation control, metabolic adaptation, and stress-responsive gene expression. Dysregulated EBF-family transcriptional circuitry has been associated with aberrant differentiation states and disease-relevant transcriptional reprogramming, making EBF4 a useful target for mechanistic studies of gene regulatory networks.

EBF4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EBF4 expression without altering the underlying DNA sequence.

EBF4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EBF4 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 EBF4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EBF4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EBF4 locus and enabling the study of EBF4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EBF4 pathway restoration in tumor cells with silenced or reduced EBF4 expression.

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