EphB3 CRISPR Activation Plasmid (h): sc-403039-ACT

EPHB3 encodes the EphB3 receptor tyrosine kinase, a member of the Eph/ephrin signaling system that mediates contact-dependent communication between neighboring cells. EphB3 participates in bidirectional signaling that coordinates cell adhesion, cytoskeletal remodeling, migration, and boundary formation during development and tissue organization. Through interactions with ephrin-B ligands, EphB3 influences pathways governing axon guidance and epithelial architecture, with downstream effects on Rho family GTPases and related phosphorylation networks. Dysregulated EphB3 signaling has been linked to altered cell positioning and invasive behavior in cancer biology and to changes in neural circuit patterning in neurodevelopmental contexts.

EphB3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EPHB3 expression without altering the underlying DNA sequence.

EphB3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EPHB3 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 EPHB3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EphB3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EPHB3 locus and enabling the study of EphB3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EphB3 pathway restoration in tumor cells with silenced or reduced EPHB3 expression.

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