CHAC1 CRISPR Activation Plasmid (m): sc-427280-ACT

Chac1 encodes CHAC1, a cytosolic γ-glutamyl cyclotransferase that degrades glutathione and modulates intracellular redox balance. CHAC1 is induced by the integrated stress response and unfolded protein response, linking ER stress signaling to amino acid availability, oxidative stress, and cell fate decisions. By lowering glutathione pools, CHAC1 can influence ROS-dependent signaling and sensitivity to lipid peroxidation pathways, including ferroptosis-associated processes. Dysregulated CHAC1 expression has been connected to cellular stress phenotypes relevant to neurodegeneration, inflammation, and cancer biology, making mouse Chac1 a useful node for mechanistic studies of stress-adaptive transcriptional programs.

CHAC1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Chac1 expression without altering the underlying DNA sequence.

CHAC1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Chac1 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 Chac1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CHAC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Chac1 locus and enabling the study of CHAC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CHAC1 pathway restoration in tumor cells with silenced or reduced Chac1 expression.

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