granzyme A CRISPR Activation Plasmid (h): sc-403958-ACT

Human GZMA encodes granzyme A, a tryptase serine protease stored in cytotoxic lymphocyte granules and released during immune synapse engagement to promote target-cell damage and inflammatory signaling. Granzyme A contributes to caspase-independent cytotoxic mechanisms and can modulate pathways linked to DNA damage responses, mitochondrial dysfunction, and cytokine production in affected cells. Its expression and activity are studied in the context of antiviral and antitumor immune surveillance, as well as dysregulated cytotoxic programs associated with autoimmunity and chronic inflammation. Because GZMA is a marker of activated cytotoxic T cells and NK cells, it is frequently used to interrogate immune-cell state transitions, effector function, and microenvironmental immune dynamics.

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

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

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