RAGE CRISPR Activation Plasmid (h): sc-400284-ACT

Human AGER encodes the receptor for advanced glycation end products (RAGE), a multiligand pattern-recognition receptor of the immunoglobulin superfamily that binds AGEs, S100/calgranulins, HMGB1, and amyloid-β. RAGE signaling amplifies inflammatory and stress responses through NF-κB, MAPK/ERK, JNK/p38, and PI3K/AKT pathways, promoting cytokine production, oxidative stress, and changes in cell adhesion and migration. It contributes to endothelial activation and leukocyte recruitment, linking metabolic and inflammatory cues to tissue remodeling. Dysregulated AGER/RAGE activity has been associated with diabetes-related vascular dysfunction, atherosclerosis, chronic inflammation, neurodegeneration, and tumor-associated inflammation, making it a common node for pathway and biomarker studies.

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

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

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