Mast Cell Tryptase CRISPR Activation Plasmid (h): sc-400887-ACT

TPSAB1 encodes mast cell tryptase (tryptase-α/β), a secreted serine protease stored in mast cell granules and released during degranulation. Mast cell tryptase participates in proteolytic remodeling of the extracellular matrix and can signal through protease-activated receptors, shaping inflammatory cascades, leukocyte recruitment, and vascular permeability. By influencing cytokine networks and tissue microenvironment dynamics, TPSAB1 is commonly used as a marker of mast cell activation and abundance in immunology and barrier tissue biology. Altered mast cell tryptase activity and mast cell infiltration patterns are studied in allergy, asthma, chronic inflammatory disorders, fibrosis-associated processes, and tumor-associated inflammation.

Mast Cell Tryptase CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TPSAB1 expression without altering the underlying DNA sequence.

Mast Cell Tryptase CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TPSAB1 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 TPSAB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Mast Cell Tryptase expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TPSAB1 locus and enabling the study of Mast Cell Tryptase-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Mast Cell Tryptase pathway restoration in tumor cells with silenced or reduced TPSAB1 expression.

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