SESN2 CRISPR Activation Plasmid (h): sc-402442-ACT

Human SESN2 (Sestrin-2) is a stress-inducible protein that coordinates cellular adaptation to metabolic, oxidative, and genotoxic insults. It modulates AMPK–mTORC1 signaling, supports redox homeostasis through antioxidant responses, and contributes to autophagy and nutrient-sensing programs that shape cell survival and growth. SESN2 is frequently studied in the context of inflammation and mitochondrial stress, where it influences ROS accumulation and downstream transcriptional networks. Dysregulated SESN2 signaling has been associated with cardiometabolic dysfunction, neurodegeneration, and cancer-related stress phenotypes, making it a useful node for pathway-level studies.

SESN2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SESN2 expression without altering the underlying DNA sequence.

SESN2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SESN2 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 SESN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SESN2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SESN2 locus and enabling the study of SESN2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SESN2 pathway restoration in tumor cells with silenced or reduced SESN2 expression.

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