Flt-1/VEGFR1 CRISPR Activation Plasmid (h): sc-400330-ACT

FLT1 encodes Flt-1/VEGFR1, a high-affinity receptor tyrosine kinase for VEGF-A, VEGF-B, and placental growth factor that modulates ligand availability and signaling output in endothelial and myeloid lineages. VEGFR1 participates in angiogenesis, vascular permeability, endothelial migration, and inflammatory cell recruitment through pathways including PI3K–AKT, MAPK/ERK, and PLCγ-dependent signaling. Both membrane-bound and soluble VEGFR1 isoforms shape VEGF gradients and vessel maturation, linking FLT1 regulation to hypoxia responses and tissue remodeling. Dysregulated FLT1/VEGFR1 signaling has been associated with pathological neovascularization and vascular dysfunction observed across cancer biology, ischemic settings, and inflammatory diseases, supporting its use as a mechanistic node in vascular and microenvironment research.

Flt-1/VEGFR1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FLT1 expression without altering the underlying DNA sequence.

Flt-1/VEGFR1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FLT1 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 FLT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Flt-1/VEGFR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FLT1 locus and enabling the study of Flt-1/VEGFR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Flt-1/VEGFR1 pathway restoration in tumor cells with silenced or reduced FLT1 expression.

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