EphB1 CRISPR Activation Plasmid (h): sc-401429-ACT

EPHB1 encodes EphB1, a receptor tyrosine kinase for ephrin-B ligands that mediates contact-dependent signaling controlling cell adhesion, repulsion, and boundary formation. EphB1 signaling engages bidirectional ephrin pathways and intersects with Rho-family GTPase regulation, MAPK/ERK cascades, and cytoskeletal remodeling to coordinate migration, axon guidance, and tissue patterning. In human biology, dysregulated Eph/ephrin signaling is frequently studied in the context of altered cell positioning, invasive behavior, and aberrant developmental programs. EPHB1 is therefore relevant to mechanistic investigations of receptor tyrosine kinase networks that shape microenvironmental interactions and phenotype switching.

EphB1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EPHB1 expression without altering the underlying DNA sequence.

EphB1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EPHB1 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 EPHB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EphB1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EPHB1 locus and enabling the study of EphB1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EphB1 pathway restoration in tumor cells with silenced or reduced EPHB1 expression.

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