neuropilin-1 CRISPR Activation Plasmid (h): sc-400428-ACT

NRP1 encodes neuropilin-1, a multifunctional transmembrane co-receptor that modulates signaling by class 3 semaphorins and vascular endothelial growth factors, shaping guidance cues, angiogenic sprouting, and tissue morphogenesis. Neuropilin-1 coordinates receptor complex formation with VEGFRs and plexins to influence cytoskeletal dynamics, migration, adhesion, and endocytosis, with downstream effects on PI3K–AKT, MAPK/ERK, and Rho GTPase pathways. In human biology, altered NRP1 expression is associated with dysregulated vascular remodeling, immune cell trafficking, and tumor microenvironment signaling, making it a widely studied node in cell–cell communication networks. These functions support research in endothelial biology, neurodevelopmental patterning, and mechanisms of invasive growth and metastasis.

neuropilin-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NRP1 expression without altering the underlying DNA sequence.

neuropilin-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NRP1 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 NRP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous neuropilin-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NRP1 locus and enabling the study of neuropilin-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of neuropilin-1 pathway restoration in tumor cells with silenced or reduced NRP1 expression.

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