cathepsin H CRISPR Activation Plasmid (h): sc-402601-ACT

CTSH encodes cathepsin H, a lysosomal cysteine protease that exhibits both endopeptidase and aminopeptidase activity, contributing to intracellular protein turnover and peptide processing. Cathepsin H participates in lysosome-dependent proteolysis and interfaces with endo-lysosomal trafficking pathways that influence antigen processing, extracellular matrix remodeling, and cellular stress responses. Altered CTSH expression or activity has been associated with dysregulated protease networks in inflammation, neurodegeneration, and cancer-related processes, where changes in lysosomal function can affect invasion, survival, and immune signaling. As a result, CTSH is frequently studied in the context of proteostasis, tumor microenvironment biology, and lysosome-centered pathway regulation.

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

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

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