ADM CRISPR Activation Plasmid (h): sc-402779-ACT

Human ADM encodes adrenomedullin, a secreted peptide hormone widely expressed in vascular and epithelial tissues that regulates vasodilation, endothelial barrier integrity, and fluid homeostasis. Through signaling via calcitonin receptor-like receptor (CALCRL) coupled to RAMP2 or RAMP3, ADM activates cAMP/PKA and downstream MAPK and PI3K–AKT pathways to modulate angiogenesis, cellular survival, and inflammatory responses. ADM is responsive to hypoxia and cellular stress and contributes to adaptive programs that shape vascular remodeling and tissue perfusion. Dysregulated ADM expression has been linked to cardiovascular dysfunction, sepsis-associated vascular leakage, and tumor-associated angiogenesis, making it a useful target for mechanistic studies of endothelial biology and microenvironmental signaling.

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

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

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