VEGF-B CRISPR Activation Plasmid (h): sc-401522-ACT

VEGFB encodes vascular endothelial growth factor B (VEGF-B), a secreted growth factor in the VEGF family that signals primarily through VEGFR1/FLT1 and the co-receptor neuropilin-1. VEGF-B is implicated in endothelial–parenchymal communication, regulation of vascular homeostasis, and lipid uptake and metabolism in energy-demanding tissues, intersecting with angiogenic and metabolic signaling programs. In cellular models it can influence survival pathways and stress responses, and its expression is often evaluated alongside hypoxia- and nutrient-sensing networks. Dysregulated VEGFB signaling has been associated in the literature with cardiometabolic phenotypes and vascular pathobiology, supporting its use as a mechanistic node in disease-relevant research systems.

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

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

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