SQSTM1/p62 CRISPR Activation Plasmid (m): sc-422075-ACT

Mouse Sqstm1 encodes SQSTM1/p62, a multifunctional adaptor that binds ubiquitinated cargo and LC3 to coordinate selective autophagy, including aggrephagy and mitophagy, while also integrating stress-response signaling. Through interactions with KEAP1, SQSTM1/p62 modulates NRF2-dependent antioxidant transcription, and it scaffolds pathways such as NF-κB and mTORC1 that link nutrient sensing to inflammation and proteostasis. Altered SQSTM1/p62 abundance or flux is frequently used as a readout of autophagic turnover and is relevant to research on protein aggregation, oxidative stress, and immune signaling in neurodegeneration, metabolic dysfunction, and cancer biology. In mouse models, Sqstm1 perturbation helps dissect how autophagy and ubiquitin signaling shape tissue homeostasis and stress adaptation.

SQSTM1/p62 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Sqstm1 expression without altering the underlying DNA sequence.

SQSTM1/p62 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Sqstm1 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 Sqstm1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SQSTM1/p62 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Sqstm1 locus and enabling the study of SQSTM1/p62-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SQSTM1/p62 pathway restoration in tumor cells with silenced or reduced Sqstm1 expression.

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