ACE2 CRISPR Activation Plasmid (h): sc-401131-ACT

Angiotensin-converting enzyme 2 (ACE2) is a human type I membrane metalloprotease that counterbalances classical renin–angiotensin system signaling by converting angiotensin II to angiotensin-(1–7), thereby shifting signaling toward MAS receptor–linked pathways. Through regulation of vasoactive peptides, ACE2 influences endothelial homeostasis, inflammatory signaling, oxidative stress responses, and tissue remodeling across cardiovascular, renal, and pulmonary systems. ACE2 also serves as a cell-surface entry factor for certain coronaviruses, linking its expression levels to studies of host–pathogen interactions and epithelial barrier biology. Dysregulated ACE2 activity or expression is frequently investigated in hypertension, cardiovascular disease, acute and chronic lung injury, kidney disease, and metabolic inflammation models.

ACE2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ACE2 expression without altering the underlying DNA sequence.

ACE2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ACE2 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 ACE2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ACE2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ACE2 locus and enabling the study of ACE2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ACE2 pathway restoration in tumor cells with silenced or reduced ACE2 expression.

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