CBS CRISPR Activation Plasmid (m): sc-419496-ACT

Mouse Cbs encodes cystathionine β-synthase (CBS), a pyridoxal phosphate–dependent enzyme that catalyzes the condensation of homocysteine and serine to form cystathionine in the transsulfuration pathway. By controlling homocysteine flux toward cysteine and glutathione biosynthesis, CBS influences cellular redox balance, methylation capacity, and sulfur amino acid homeostasis, with downstream effects on oxidative stress and mitochondrial function. CBS also contributes to endogenous hydrogen sulfide production, linking Cbs activity to signaling processes that modulate vascular tone and inflammatory responses. Dysregulation of CBS-dependent metabolism is widely used to model hyperhomocysteinemia-associated pathophysiology and to interrogate metabolic contributions to neurovascular and hepatic phenotypes in mouse systems.

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

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

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