TPPII CRISPR Activation Plasmid (h): sc-403717-ACT

Human TPP2 encodes tripeptidyl peptidase II (TPPII), a large cytosolic serine exopeptidase that trims oligopeptides generated by the ubiquitin–proteasome system and supports protein quality control and intracellular peptide turnover. By regulating peptide processing, TPPII influences proteostasis, antigenic peptide availability for MHC class I presentation, and stress-adaptive pathways linked to cell survival and apoptosis. Altered TPP2/TPPII activity has been associated with dysregulated proteolysis and immune-related phenotypes, making it relevant for mechanistic studies of proteasome compensation, inflammatory signaling, and cell-state transitions under proteotoxic stress. TPP2 is therefore a useful node for dissecting how cytosolic peptidases shape protein degradation pathways and downstream cellular responses in human model systems.

TPPII CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TPP2 expression without altering the underlying DNA sequence.

TPPII CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TPP2 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 TPP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TPPII expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TPP2 locus and enabling the study of TPPII-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TPPII pathway restoration in tumor cells with silenced or reduced TPP2 expression.

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