p53 CRISPR Activation Plasmid (m): sc-423509-ACT

Mouse Trp53 encodes the p53 transcription factor, a central regulator of genome integrity that integrates DNA damage, oncogenic stress, and metabolic cues to coordinate cell-cycle arrest, senescence, apoptosis, and DNA repair. p53 activity intersects with ATM/ATR–CHK signaling, the MDM2 negative-feedback loop, and pathways controlling oxidative stress and mitochondrial homeostasis. Disruption or attenuation of p53 signaling is broadly relevant to tumor biology and influences experimental outcomes in studies of genotoxic stress, chromatin regulation, and immune-modulatory gene programs. In murine model systems, tuning Trp53 expression is commonly used to probe stress-adaptation mechanisms and transformation-associated phenotypes.

p53 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Trp53 expression without altering the underlying DNA sequence.

p53 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Trp53 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 Trp53 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p53 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Trp53 locus and enabling the study of p53-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p53 pathway restoration in tumor cells with silenced or reduced Trp53 expression.

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