ATG7 CRISPR Activation Plasmid (m): sc-428805-ACT

Mouse Atg7 encodes ATG7, an E1-like enzyme that catalyzes ubiquitin-like conjugation reactions essential for autophagosome formation. ATG7 activates ATG12 and LC3/Atg8 family proteins to drive phagophore expansion, cargo sequestration, and autophagic flux, linking nutrient sensing to lysosomal turnover. Through its central role in macroautophagy, ATG7 influences mitochondrial quality control, proteostasis, and innate immune signaling, and its dysregulation is commonly used to model autophagy-dependent phenotypes in neurodegeneration, metabolic stress, infection biology, and cancer-relevant cellular adaptation. In mouse systems, Atg7 perturbation is widely applied to interrogate pathway crosstalk with mTOR/AMPK signaling, ER stress responses, and inflammation-associated transcriptional programs.

ATG7 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Atg7 expression without altering the underlying DNA sequence.

ATG7 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Atg7 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 Atg7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ATG7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Atg7 locus and enabling the study of ATG7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ATG7 pathway restoration in tumor cells with silenced or reduced Atg7 expression.

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