ephrin-A5 CRISPR Activation Plasmid (m): sc-420123-ACT

Efna5 encodes ephrin-A5, a glycosylphosphatidylinositol-anchored ligand for EphA receptors that mediates contact-dependent cell signaling controlling axon guidance, neuronal migration, and synaptic organization. Ephrin-A5–Eph forward and reverse signaling coordinates cytoskeletal remodeling through Rho-family GTPases and intersects with MAPK/ERK and PI3K-associated pathways to regulate adhesion, repulsion, and tissue boundary formation. In mouse systems, Efna5 activity is widely used to model mechanisms of neural circuit assembly, retinal mapping, and developmental patterning. Dysregulated ephrin/Eph signaling is implicated in neurodevelopmental phenotypes and aberrant cell motility programs relevant to cancer biology and angiogenesis research contexts.

ephrin-A5 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Efna5 expression without altering the underlying DNA sequence.

ephrin-A5 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Efna5 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 Efna5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ephrin-A5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Efna5 locus and enabling the study of ephrin-A5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ephrin-A5 pathway restoration in tumor cells with silenced or reduced Efna5 expression.

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