EVL CRISPR Activation Plasmid (h): sc-404876-ACT

Human EVL (Ena/VASP-like) encodes an actin regulatory protein of the Ena/VASP family that promotes filament elongation and controls lamellipodia and filopodia dynamics during cell spreading, adhesion, and migration. EVL functions downstream of signaling from integrins and receptor tyrosine kinases, coordinating cytoskeletal remodeling with focal adhesion turnover and membrane protrusion. By shaping actin architecture, EVL contributes to processes such as immune cell trafficking, endothelial barrier regulation, and neuronal outgrowth. Altered EVL expression or pathway context has been studied in disorders involving dysregulated cell motility and tissue remodeling, including invasive cancer phenotypes and inflammatory pathobiology.

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

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

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