GSDMDC1 CRISPR Activation Plasmid (h): sc-406596-ACT

GSDMD (gasdermin D; also referenced as GSDMDC1) encodes a pore-forming effector of inflammatory cell death that is activated by proteolytic cleavage downstream of inflammasome signaling. Upon cleavage by caspase-1 or caspase-4/5/11 pathways, the N-terminal fragment oligomerizes in membranes to drive pyroptosis and promote release of IL-1 family cytokines, linking innate immune sensing to lytic cell death. This axis integrates with NLRP3 and related inflammasome pathways, shaping antimicrobial defense and sterile inflammation. Dysregulated GSDMD activity has been implicated in chronic inflammatory states, sepsis-associated inflammation models, and tumor–immune interactions, motivating mechanistic studies of cell death and cytokine biology in human systems.

GSDMDC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GSDMD expression without altering the underlying DNA sequence.

GSDMDC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GSDMD 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 GSDMD transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GSDMDC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GSDMD locus and enabling the study of GSDMDC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GSDMDC1 pathway restoration in tumor cells with silenced or reduced GSDMD expression.

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