ALK-1 CRISPR Activation Plasmid (h): sc-400728-ACT

ACVRL1 encodes activin receptor-like kinase 1 (ALK-1), an endothelial-enriched type I serine/threonine kinase receptor in the TGF-β superfamily that transduces BMP9/BMP10 signals. Upon ligand engagement, ALK-1 cooperates with type II receptors to phosphorylate SMAD1/5/8 and coordinate transcriptional programs governing angiogenesis, vascular remodeling, and endothelial quiescence. This signaling axis intersects with TGF-β/SMAD2/3 balance and Notch-dependent cues to regulate vessel maturation and arteriovenous identity. Dysregulated ACVRL1 activity is linked to heritable vascular malformations, including phenotypes associated with hereditary hemorrhagic telangiectasia, making it a key target for mechanistic studies in vascular biology.

ALK-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ACVRL1 expression without altering the underlying DNA sequence.

ALK-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ACVRL1 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 ACVRL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ALK-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ACVRL1 locus and enabling the study of ALK-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ALK-1 pathway restoration in tumor cells with silenced or reduced ACVRL1 expression.

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