p300 CRISPR Activation Plasmid (h): sc-400055-ACT

Human EP300 encodes p300, a lysine acetyltransferase and transcriptional coactivator that integrates signals from multiple transcription factors to regulate chromatin accessibility and gene expression. p300 acetylates histones and non-histone proteins, shaping enhancer activity and controlling programs involved in cell cycle progression, differentiation, DNA damage responses, and stress signaling. EP300 participates in pathways linked to MAPK/ERK, TGF-β/SMAD, hypoxia (HIF), and p53-mediated transcriptional regulation through broad protein–protein interaction networks. Dysregulation of EP300/p300-dependent acetylation and transcriptional control is associated with altered lineage specification and aberrant transcriptional states observed across diverse disease models.

p300 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EP300 expression without altering the underlying DNA sequence.

p300 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EP300 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 EP300 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p300 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EP300 locus and enabling the study of p300-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p300 pathway restoration in tumor cells with silenced or reduced EP300 expression.

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