gasdermin CRISPR Activation Plasmid (h): sc-410658-ACT

Human GSDMA encodes gasdermin A, a pore-forming effector of regulated cell death that contributes to epithelial barrier biology and inflammatory signaling. Gasdermins can be activated by proteolytic processing to unmask an N-terminal domain that oligomerizes in membranes, promoting lytic death programs and release of pro-inflammatory mediators. GSDMA expression and regulation are linked to stress-response pathways in stratified epithelia, with reported associations to asthma, inflammatory skin phenotypes, and susceptibility loci in immune-mediated disease. As a result, GSDMA is frequently studied in the context of epithelial homeostasis, innate immune responses, and cell death–driven tissue remodeling.

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

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

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