ENDOG CRISPR Activation Plasmid (h): sc-403263-ACT

Human ENDOG (endonuclease G) is a mitochondrially localized nuclease implicated in mitochondrial DNA maintenance and the execution of caspase-independent cell death programs. Upon mitochondrial outer membrane permeabilization, ENDOG can translocate to the nucleus and contribute to large-scale chromatin degradation, linking mitochondrial stress signaling to genome integrity outcomes. It also intersects with oxidative stress responses and bioenergetic homeostasis, processes frequently perturbed in neurodegeneration, ischemic injury models, and cancer-associated mitochondrial dysfunction. Dysregulated ENDOG activity has been examined in contexts of altered apoptosis susceptibility, mitochondrial genome instability, and inflammatory signaling driven by damaged DNA.

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

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

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