EGFL7 CRISPR Activation Plasmid (m): sc-436135-ACT

Egfl7 encodes epidermal growth factor-like domain 7 (EGFL7), a secreted extracellular matrix–associated protein enriched in endothelial cells that supports vascular development and angiogenic remodeling. EGFL7 participates in endothelial migration, tube formation, and vessel stabilization, and has been linked to modulation of cell–cell and cell–matrix signaling, including effects on Notch pathway activity during sprouting angiogenesis. In mouse models, altered Egfl7 expression perturbs vascular patterning and endothelial behavior, implicating the gene in pathways relevant to ischemic injury responses and tumor-associated neovascularization. Dysregulated EGFL7 has also been studied in the context of inflammatory microenvironments and tissue remodeling where endothelial dysfunction contributes to disease phenotypes.

EGFL7 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Egfl7 expression without altering the underlying DNA sequence.

EGFL7 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Egfl7 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 Egfl7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EGFL7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Egfl7 locus and enabling the study of EGFL7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EGFL7 pathway restoration in tumor cells with silenced or reduced Egfl7 expression.

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