ML-IAP CRISPR Activation Plasmid (h): sc-403976-ACT

BIRC7 encodes ML-IAP (also known as livin), a member of the inhibitor of apoptosis (IAP) protein family that suppresses programmed cell death by binding and inhibiting effector caspases such as CASP3, CASP7, and CASP9. Through its BIR and RING domains, ML-IAP can modulate apoptosis signaling and ubiquitin-dependent protein turnover, shaping cellular responses to intrinsic and extrinsic death cues. Dysregulated BIRC7 expression has been associated with apoptosis resistance and altered stress signaling in multiple cancer contexts, supporting its use as a mechanistic node in survival pathway studies. In research settings, ML-IAP is frequently examined alongside TNF signaling, mitochondrial apoptosis regulation, and cellular proteostasis networks.

ML-IAP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BIRC7 expression without altering the underlying DNA sequence.

ML-IAP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BIRC7 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 BIRC7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ML-IAP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BIRC7 locus and enabling the study of ML-IAP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ML-IAP pathway restoration in tumor cells with silenced or reduced BIRC7 expression.

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