c-IAP2 CRISPR Activation Plasmid (h): sc-400762-ACT

Human BIRC3 encodes c-IAP2, a RING-domain E3 ubiquitin ligase that regulates apoptosis and inflammatory signaling by ubiquitinating key components of TNF receptor complexes and modulating caspase activation. c-IAP2 participates in NF-κB pathway control, including crosstalk between canonical and non-canonical NF-κB signaling through regulation of NIK stability and ubiquitin-dependent checkpointing of receptor-proximal signaling. Through these functions, BIRC3 influences cell survival, cytokine-induced responses, and stress adaptation across immune and epithelial contexts. Dysregulation of BIRC3 expression or activity has been linked to altered apoptotic thresholds and aberrant NF-κB signaling observed in multiple disease-relevant models, supporting its use as a mechanistic node in cell death and inflammation research.

c-IAP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BIRC3 expression without altering the underlying DNA sequence.

c-IAP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BIRC3 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 BIRC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous c-IAP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BIRC3 locus and enabling the study of c-IAP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of c-IAP2 pathway restoration in tumor cells with silenced or reduced BIRC3 expression.

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