TMPRSS11A CRISPR Activation Plasmid (h): sc-415530-ACT

TMPRSS11A encodes a type II transmembrane serine protease that localizes to the cell surface and participates in pericellular proteolysis at epithelial interfaces. By cleaving extracellular and membrane-associated substrates, TMPRSS11A can influence epithelial differentiation programs, barrier maintenance, and protease-activated signaling networks that shape tissue remodeling and inflammatory responses. Its activity intersects with broader serine protease cascades and can modulate the availability of bioactive peptides and growth-factor–related cues in the local microenvironment. Dysregulated expression of membrane-anchored serine proteases, including TMPRSS11A, has been associated with altered airway and mucosal biology and has been explored in the context of epithelial pathophysiology and cancer-related protease remodeling.

TMPRSS11A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMPRSS11A expression without altering the underlying DNA sequence.

TMPRSS11A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMPRSS11A 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 TMPRSS11A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMPRSS11A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMPRSS11A locus and enabling the study of TMPRSS11A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMPRSS11A pathway restoration in tumor cells with silenced or reduced TMPRSS11A expression.

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