VEGF-C CRISPR Activation Plasmid (m): sc-423667-ACT

Mouse Vegfc encodes vascular endothelial growth factor C (VEGF-C), a secreted ligand that primarily signals through VEGFR-3/FLT4 to regulate lymphangiogenesis and lymphatic vessel remodeling, with additional effects on angiogenic programs in certain contexts. VEGF-C influences endothelial proliferation, migration, and survival, integrating with downstream MAPK/ERK and PI3K/AKT signaling to shape vascular network formation and permeability. In immune and stromal microenvironments, VEGF-C–driven lymphatic changes can modulate interstitial fluid balance and trafficking of immune cells via lymphatic conduits. Dysregulated VEGF-C/VEGFR-3 signaling is linked to pathological lymphatic remodeling and is frequently studied in models of inflammation, tissue fibrosis, and tumor-associated lymphangiogenesis.

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

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

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