TP53INP2 CRISPR Activation Plasmid (h): sc-405261-ACT

TP53INP2 (tumor protein p53 inducible nuclear protein 2) is a stress-responsive protein implicated in autophagy regulation and intracellular trafficking, with reported roles in organizing autophagosome biogenesis and supporting selective autophagy programs. It has been linked to cytoplasmic-to-nuclear shuttling and interactions with autophagy machinery, connecting p53-associated stress responses to catabolic remodeling. Through these functions, TP53INP2 can influence cellular homeostasis under nutrient deprivation and other stressors, impacting pathways that govern survival, metabolism, and quality control. Dysregulated autophagy and stress signaling involving TP53INP2 are relevant to mechanistic studies in cancer biology, neurodegeneration, and inflammatory phenotypes where proteostasis and metabolic adaptation are perturbed.

TP53INP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TP53INP2 expression without altering the underlying DNA sequence.

TP53INP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TP53INP2 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 TP53INP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TP53INP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TP53INP2 locus and enabling the study of TP53INP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TP53INP2 pathway restoration in tumor cells with silenced or reduced TP53INP2 expression.

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