xCT CRISPR Activation Plasmid (h): sc-401920-ACT

Human SLC7A11 encodes xCT, the light chain subunit of system xC⁻ that mediates sodium-independent cystine import in exchange for intracellular glutamate. By supplying cystine for glutathione synthesis, xCT supports cellular redox homeostasis, detoxification of reactive oxygen species, and metabolic coupling between amino acid transport and antioxidant capacity. SLC7A11 is regulated by stress-responsive programs such as NRF2 and integrated stress signaling, linking it to ferroptosis sensitivity and broader thiol metabolism. Dysregulated xCT activity has been associated with altered oxidative stress handling and metabolic rewiring observed across multiple disease-relevant contexts, including tumor biology and neuroinflammatory processes.

xCT CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC7A11 expression without altering the underlying DNA sequence.

xCT CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC7A11 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 SLC7A11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous xCT expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC7A11 locus and enabling the study of xCT-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of xCT pathway restoration in tumor cells with silenced or reduced SLC7A11 expression.

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