EBP1 CRISPR Activation Plasmid (h): sc-402177-ACT

PA2G4 encodes ErbB3-binding protein 1 (EBP1), a multifunctional RNA- and protein-binding factor implicated in coordinating transcriptional and translational programs that influence cell-cycle progression and cellular differentiation. EBP1 associates with ErbB receptor signaling and participates in regulation of gene expression through interactions with chromatin- and ribonucleoprotein-linked processes, impacting pathways that control proliferation and stress responses. Altered PA2G4/EBP1 expression or activity has been studied in the context of oncogenic signaling networks, including changes in growth factor–responsive transcriptional outputs. These properties make PA2G4 a useful node for probing how receptor-driven cues couple to gene-expression control in human cells.

EBP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PA2G4 expression without altering the underlying DNA sequence.

EBP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PA2G4 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 PA2G4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EBP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PA2G4 locus and enabling the study of EBP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EBP1 pathway restoration in tumor cells with silenced or reduced PA2G4 expression.

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