EGFL6 CRISPR Activation Plasmid (h): sc-411850-ACT

EGFL6 (epidermal growth factor-like protein 6) is a secreted extracellular matrix–associated protein enriched in vascular and stromal compartments, where it supports cell–matrix interactions and communication within the tissue microenvironment. Through its EGF-like repeats, EGFL6 influences endothelial and mesenchymal cell behaviors linked to adhesion, migration, and angiogenic remodeling, processes that intersect with integrin signaling and growth factor–responsive pathways. Altered EGFL6 expression has been reported in multiple tumor contexts and other disorders characterized by aberrant neovascularization and extracellular matrix reprogramming, making it a useful marker and functional node for studying microenvironment-driven phenotypes. Human EGFL6 research commonly focuses on how extracellular cues reshape transcriptional programs that control invasion, vascular patterning, and tissue organization.

EGFL6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EGFL6 expression without altering the underlying DNA sequence.

EGFL6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EGFL6 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 EGFL6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EGFL6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EGFL6 locus and enabling the study of EGFL6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EGFL6 pathway restoration in tumor cells with silenced or reduced EGFL6 expression.

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