Atg13 CRISPR Activation Plasmid (h2): sc-404702-ACT-2

Human ATG13 encodes Atg13, a core component of the ULK1 initiation complex that integrates nutrient and energy signals from mTORC1 and AMPK to trigger autophagosome formation. By coordinating early autophagy events and phagophore nucleation through interactions with ULK1/2, FIP200/RB1CC1, and ATG101, Atg13 supports proteostasis, organelle quality control, and stress adaptation. Dysregulated ATG13-dependent autophagy has been implicated in pathways relevant to cancer biology, neurodegeneration, and inflammatory signaling, where altered autophagic flux can impact cell survival and innate immune responses. Gene editing of ATG13 enables mechanistic studies of autophagy initiation, mapping of signaling crosstalk, and functional genomics screens that quantify autophagic flux, stress sensitivity, and pathway dependencies in human cellular models.

Atg13 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous ATG13 expression without altering the underlying DNA sequence.

Atg13 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ATG13 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 ATG13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Atg13 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ATG13 locus and enabling the study of Atg13-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Atg13 pathway restoration in tumor cells with silenced or reduced ATG13 expression.

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