ephrin-A2 CRISPR Activation Plasmid (h): sc-403672-ACT

EFNA2 encodes ephrin-A2, a glycosylphosphatidylinositol-anchored membrane ligand for Eph receptor tyrosine kinases that mediates contact-dependent signaling in cell–cell communication. Eph/ephrin signaling regulates axon guidance, neuronal circuit assembly, vascular patterning, epithelial boundary formation, and cell migration through pathways that interface with Rho-family GTPases, MAPK signaling, and cytoskeletal remodeling. Altered EFNA2 expression or EphA–ephrin-A signaling balance has been associated with dysregulated adhesion and motility phenotypes relevant to neurodevelopmental processes and cancer biology, including tumor invasion and angiogenic remodeling. As a surface-tethered cue, ephrin-A2 is also useful for studying bidirectional signaling, where ligand- and receptor-expressing cells trigger distinct downstream responses.

ephrin-A2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EFNA2 expression without altering the underlying DNA sequence.

ephrin-A2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EFNA2 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 EFNA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ephrin-A2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EFNA2 locus and enabling the study of ephrin-A2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ephrin-A2 pathway restoration in tumor cells with silenced or reduced EFNA2 expression.

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