p300 CRISPR Activation Plasmid (m): sc-435902-ACT

Mouse Ep300 encodes p300, a broadly acting transcriptional coactivator and lysine acetyltransferase that acetylates histones and diverse non-histone substrates to regulate chromatin accessibility and gene expression programs. p300 integrates signaling inputs from pathways such as TGF-β/SMAD, Wnt/β-catenin, nuclear receptor signaling, and hypoxia responses through protein–protein interactions and enhancer binding, shaping cell fate decisions, proliferation, and stress responses. Ep300 activity influences enhancer and super-enhancer function, DNA damage responses, and inflammatory transcriptional networks, making it central to epigenetic control of development and tissue homeostasis. Dysregulated p300-dependent acetylation and coactivation is associated with altered differentiation and transcriptional rewiring observed across multiple disease-relevant biological contexts, including cancer models and inflammatory states.

p300 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ep300 expression without altering the underlying DNA sequence.

p300 CRISPR Activation Plasmid (m) 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.