p73 CRISPR Activation Plasmid (h): sc-416822-ACT

Human TP73 encodes p73, a p53 family transcription factor that regulates cell fate decisions including cell cycle arrest, apoptosis, differentiation, and senescence. Through context-dependent transactivation of target genes, p73 integrates stress responses and influences DNA damage signaling, mitochondrial apoptosis programs, and developmental pathways. Isoform diversity (TAp73 and ΔNp73) can shift transcriptional outputs toward pro-apoptotic or pro-survival states, shaping epithelial integrity and neuronal development. Dysregulated TP73 activity and altered isoform balance are frequently studied in cancer biology, neurodevelopmental disorders, and chemoresistance mechanisms as they modulate transcriptional networks and genomic stability.

p73 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TP73 expression without altering the underlying DNA sequence.

p73 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TP73 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 TP73 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p73 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TP73 locus and enabling the study of p73-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p73 pathway restoration in tumor cells with silenced or reduced TP73 expression.

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