NOXA CRISPR Activation Plasmid (h): sc-400498-ACT

PMAIP1 encodes NOXA, a BH3-only pro-apoptotic member of the BCL2 family that promotes mitochondrial outer membrane permeabilization by preferentially antagonizing MCL1 and related anti-apoptotic proteins. NOXA is rapidly induced by cellular stress signals, including p53-dependent DNA damage responses, and integrates apoptosis with proteostasis and hypoxia-associated signaling. By shifting the balance of BCL2-family interactions, NOXA influences caspase activation, mitochondrial integrity, and cell fate decisions. Dysregulated PMAIP1/NOXA expression has been linked to altered apoptotic priming in cancer biology and to stress-response phenotypes relevant to neurodegeneration and inflammatory contexts.

NOXA CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PMAIP1 expression without altering the underlying DNA sequence.

NOXA CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PMAIP1 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 PMAIP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NOXA expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PMAIP1 locus and enabling the study of NOXA-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NOXA pathway restoration in tumor cells with silenced or reduced PMAIP1 expression.

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