Renin CRISPR Activation Plasmid (h): sc-400890-ACT

Human REN encodes renin, an aspartyl protease that initiates the renin–angiotensin–aldosterone system by cleaving angiotensinogen to generate angiotensin I, thereby regulating angiotensin II production and downstream signaling that controls vascular tone and electrolyte balance. Renin activity is integrated with endocrine feedback loops involving aldosterone, sympathetic signaling, and renal sodium handling, linking renal physiology to systemic hemodynamics. Dysregulated REN expression or renin pathway activation is implicated in hypertension, cardiovascular remodeling, and chronic kidney disease–associated pathophysiology. As a secreted enzyme produced primarily by juxtaglomerular cells, renin also serves as a tractable node for studying stimulus-secretion coupling and transcriptional control in renal and vascular cell models.

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

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

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