EphB2 CRISPR Activation Plasmid (h): sc-401642-ACT

EPHB2 encodes the human EphB2 receptor tyrosine kinase, a member of the Eph/ephrin signaling system that mediates contact-dependent communication controlling cell positioning, boundary formation, and axon guidance. EphB2 activation by ephrin-B ligands regulates cytoskeletal remodeling, adhesion dynamics, and migration through pathways involving Rho family GTPases, MAPK signaling, and PI3K-related networks. In mammalian tissues, EphB2 contributes to developmental patterning and maintenance of epithelial architecture, and dysregulated EphB2 signaling has been implicated in altered invasiveness and differentiation states in cancer biology. EPHB2 is also studied in the context of neurodevelopmental processes and synaptic organization where graded receptor–ligand interactions influence circuit formation.

EphB2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EPHB2 expression without altering the underlying DNA sequence.

EphB2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EPHB2 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 EPHB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EphB2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EPHB2 locus and enabling the study of EphB2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EphB2 pathway restoration in tumor cells with silenced or reduced EPHB2 expression.

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