Endoglin/CD105 CRISPR Activation Plasmid (h): sc-400599-ACT

ENG encodes endoglin (CD105), a homodimeric transmembrane glycoprotein that functions as an auxiliary receptor within the TGF-β superfamily receptor complex. It modulates SMAD-dependent signaling downstream of ligands such as TGF-β and BMPs, shaping endothelial cell proliferation, migration, and extracellular matrix remodeling during angiogenesis and vascular homeostasis. Endoglin is highly expressed on activated endothelium and certain stromal compartments, linking it to processes including vascular development, inflammation-associated remodeling, and hypoxia-responsive signaling. Dysregulated ENG expression or function is implicated in vascular malformation biology and aberrant neovascularization observed across multiple disease contexts.

Endoglin/CD105 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ENG expression without altering the underlying DNA sequence.

Endoglin/CD105 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ENG 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 ENG transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Endoglin/CD105 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ENG locus and enabling the study of Endoglin/CD105-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Endoglin/CD105 pathway restoration in tumor cells with silenced or reduced ENG expression.

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