EphA5 CRISPR Activation Plasmid (h): sc-403104-ACT

EPHA5 encodes ephrin type-A receptor 5 (EphA5), a receptor tyrosine kinase that mediates contact-dependent signaling through binding to membrane-tethered ephrin-A ligands. EphA5 regulates axon guidance, synapse formation, and neuronal circuit refinement by coordinating cytoskeletal remodeling, cell adhesion, and directional cell repulsion/attraction programs. Downstream signaling intersects with Rho-family GTPases, MAPK pathways, and kinase/phosphatase networks that control neurite outgrowth and cell migration. Dysregulated Eph/ephrin signaling has been implicated in neurodevelopmental and neuropsychiatric phenotypes as well as altered invasive behavior and pathway rewiring in cancer-related contexts.

EphA5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EPHA5 expression without altering the underlying DNA sequence.

EphA5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EPHA5 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 EPHA5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EphA5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EPHA5 locus and enabling the study of EphA5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EphA5 pathway restoration in tumor cells with silenced or reduced EPHA5 expression.

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