EBF3 CRISPR Activation Plasmid (h): sc-402181-ACT

EBF3 (early B-cell factor 3) is a helix–loop–helix transcription factor that binds DNA to regulate gene expression programs controlling neuronal differentiation, maturation, and broader developmental patterning. In human cells, EBF3 contributes to lineage-specifying transcriptional networks and cooperates with chromatin and cofactor assemblies to shape cell-state transitions and tissue-specific gene regulation. Dysregulated EBF3 expression or function has been linked to neurodevelopmental phenotypes and altered transcriptional circuitry observed in diverse disease contexts, making it a useful node for interrogating developmental gene regulatory networks. Studying EBF3-dependent pathways can clarify how transcriptional timing and promoter/enhancer selection influence cell fate decisions and stress-responsive transcription.

EBF3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EBF3 expression without altering the underlying DNA sequence.

EBF3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EBF3 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 EBF3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EBF3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EBF3 locus and enabling the study of EBF3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EBF3 pathway restoration in tumor cells with silenced or reduced EBF3 expression.

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