Apelin CRISPR Activation Plasmid (h): sc-401548-ACT

APLN encodes apelin, a secreted peptide ligand for the APJ receptor (APLNR) that regulates vascular tone, angiogenesis, and fluid homeostasis through G protein–coupled signaling. Apelin–APJ activation engages pathways such as PI3K/AKT, ERK/MAPK, and eNOS/NO, influencing endothelial function, cardiac contractility, and metabolic adaptation in multiple tissues. Dysregulation of APLN expression has been linked to cardiovascular remodeling, pulmonary vascular disease, and altered energy balance in obesity and diabetes-related contexts. In tumor biology, apelin signaling can modulate hypoxia responses and neovascularization, making it relevant for studies of microenvironmental control of growth and perfusion.

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

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

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