EphB6 CRISPR Activation Plasmid (m): sc-420202-ACT

Mouse Ephb6 encodes EphB6, a kinase-impaired member of the Eph receptor tyrosine kinase family that modulates ephrin-mediated cell–cell signaling. EphB6 participates in pathways controlling contact-dependent adhesion, cytoskeletal remodeling, and directional cell migration, and can influence downstream Rho family GTPase and MAPK signaling through receptor clustering and co-receptor interactions. In developing and adult tissues, EphB6 contributes to tissue boundary formation and immune cell communication, processes frequently implicated in altered tissue organization and dysregulated inflammatory responses. Changes in Eph/ephrin signaling are broadly associated with aberrant motility and microenvironmental interactions, making EphB6 regulation relevant for mechanistic studies in development, neurobiology, and cancer biology models.

EphB6 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ephb6 expression without altering the underlying DNA sequence.

EphB6 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ephb6 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 Ephb6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EphB6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ephb6 locus and enabling the study of EphB6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EphB6 pathway restoration in tumor cells with silenced or reduced Ephb6 expression.

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