p8 CRISPR Activation Plasmid (h): sc-402153-ACT

Human NUPR1 encodes the stress-inducible chromatin-associated protein p8, a small, intrinsically disordered regulator of transcriptional programs that coordinate adaptation to cellular injury. p8 integrates signals from ER stress, oxidative stress, and nutrient limitation to influence cell-cycle control, apoptosis, autophagy, and DNA damage responses through context-dependent modulation of transcription factor networks. It has been linked to remodeling of inflammatory and metabolic pathways and to changes in epithelial–mesenchymal traits, supporting roles in cellular plasticity. Dysregulated NUPR1/p8 activity is frequently studied in oncology and inflammation-associated pathobiology, where altered stress tolerance and survival signaling contribute to disease-relevant phenotypes.

p8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NUPR1 expression without altering the underlying DNA sequence.

p8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NUPR1 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 NUPR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NUPR1 locus and enabling the study of p8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p8 pathway restoration in tumor cells with silenced or reduced NUPR1 expression.

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