TMPRSS2 CRISPR Activation Plasmid (h): sc-402828-ACT

TMPRSS2 (transmembrane serine protease 2) encodes a type II transmembrane serine protease expressed prominently in prostate and airway epithelium, where it contributes to pericellular proteolysis and epithelial homeostasis. Its transcription is regulated by androgen receptor signaling, linking TMPRSS2 expression to hormone-responsive gene networks and epithelial differentiation programs. TMPRSS2 is widely studied for its role in protease-mediated processing of cell-surface and secreted substrates and for modulating pathways involved in tissue remodeling and inflammatory responses. Dysregulated TMPRSS2 expression and TMPRSS2-associated gene fusions are relevant to prostate cancer biology, and TMPRSS2-dependent proteolytic activity is implicated in host–pathogen interactions in respiratory research models.

TMPRSS2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMPRSS2 expression without altering the underlying DNA sequence.

TMPRSS2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMPRSS2 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 TMPRSS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMPRSS2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMPRSS2 locus and enabling the study of TMPRSS2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMPRSS2 pathway restoration in tumor cells with silenced or reduced TMPRSS2 expression.

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