XIAP CRISPR Activation Plasmid (h): sc-416497-ACT

X-linked inhibitor of apoptosis protein (XIAP) is a potent suppressor of programmed cell death that directly binds and inhibits effector caspases such as CASP3 and CASP7 as well as initiator caspase CASP9 via its BIR domains. Beyond caspase blockade, XIAP participates in ubiquitin-dependent signaling through its RING E3 ligase activity, intersecting with TNF receptor–associated pathways and modulating NF-κB–linked survival programs. Through these activities, XIAP helps determine cellular sensitivity to intrinsic and extrinsic apoptotic cues and shapes stress-adaptive responses. Dysregulated XIAP expression or function has been associated with altered apoptosis thresholds and inflammatory signaling states relevant to cancer biology and immune dysregulation research.

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

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

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