Vasohibin CRISPR Activation Plasmid (h): sc-403526-ACT

Human VASH1 encodes vasohibin-1, an endothelium-associated regulator of angiogenesis that is induced by pro-angiogenic cues and functions as a negative feedback modulator of endothelial proliferation and migration. Vasohibin-1 influences vascular remodeling and vessel maturation programs and is linked to cellular processes including extracellular matrix interactions and stress-responsive signaling in the vascular microenvironment. Altered VASH1 expression has been associated with pathological neovascularization and tumor angiogenesis, making it a useful molecular node for studying endothelial homeostasis and microvascular phenotypes. As a vascular regulatory factor, VASH1 is frequently examined in models of ischemia, inflammation-associated vascular change, and cancer biology to dissect context-dependent angiogenic signaling.

Vasohibin-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous VASH1 expression without altering the underlying DNA sequence.

Vasohibin-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the VASH1 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 VASH1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Vasohibin-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native VASH1 locus and enabling the study of Vasohibin-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Vasohibin-1 pathway restoration in tumor cells with silenced or reduced VASH1 expression.

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