ASC/TMS1/PYCARD CRISPR Activation Plasmid (h): sc-400151-ACT

PYCARD encodes ASC (also known as TMS1), a cytosolic adaptor protein that nucleates inflammasome assembly and links upstream pattern-recognition receptors to activation of inflammatory caspases. Through its PYD and CARD domains, ASC promotes formation of ASC specks, facilitating caspase-1 activation and downstream processing of IL-1β and IL-18, as well as gasdermin D–dependent pyroptosis. ASC-dependent signaling integrates with innate immune pathways including NLRP3, AIM2, and related inflammasome platforms, shaping antimicrobial defense and sterile inflammation. Dysregulated PYCARD/ASC activity has been implicated in inflammatory and autoimmune disease mechanisms and is frequently studied in contexts of infection, neuroinflammation, and tumor-associated immune signaling.

ASC/TMS1/PYCARD CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PYCARD expression without altering the underlying DNA sequence.

ASC/TMS1/PYCARD CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PYCARD 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 PYCARD transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ASC/TMS1/PYCARD expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PYCARD locus and enabling the study of ASC/TMS1/PYCARD-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ASC/TMS1/PYCARD pathway restoration in tumor cells with silenced or reduced PYCARD expression.

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