Angiotensinogen CRISPR Activation Plasmid (h): sc-401013-ACT

Human AGT encodes angiotensinogen, a secreted liver-derived glycoprotein that is the obligate precursor of angiotensin peptides in the renin–angiotensin system (RAS). Proteolytic processing of angiotensinogen initiates cascades that regulate vascular tone, fluid and electrolyte balance, and tissue remodeling through downstream angiotensin signaling networks. Beyond systemic endocrine signaling, locally produced AGT supports paracrine RAS activity in kidney, vasculature, adipose, and brain, linking it to oxidative stress, inflammation, and extracellular matrix turnover pathways. Dysregulated AGT expression and RAS imbalance are associated with cardiometabolic and renal disease biology, providing a mechanistic handle for studying hemodynamic regulation and organ crosstalk in human cell models.

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

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

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