EBF2 CRISPR Activation Plasmid (h): sc-402456-ACT

EBF2 (early B-cell factor 2) is a helix–loop–helix transcription factor that binds specific DNA motifs to regulate lineage specification and differentiation programs in human development. It plays prominent roles in adipocyte biology, neuronal differentiation, and osteogenic pathways by coordinating transcriptional networks that control metabolic gene expression and cell fate decisions. EBF2 activity integrates with chromatin remodeling and enhancer selection to maintain stable transcriptional states, and its dysregulation has been associated with altered adipose thermogenic programs, neurodevelopmental phenotypes, and cancer-related transcriptional reprogramming. As a nodal regulator of tissue-specific gene expression, EBF2 is frequently studied in the context of epigenetic control, metabolic homeostasis, and differentiation-linked signaling cascades.

EBF2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EBF2 expression without altering the underlying DNA sequence.

EBF2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EBF2 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 EBF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EBF2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EBF2 locus and enabling the study of EBF2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EBF2 pathway restoration in tumor cells with silenced or reduced EBF2 expression.

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