EphA7 CRISPR Activation Plasmid (h): sc-401722-ACT

EPHA7 encodes EphA7, a member of the ephrin receptor tyrosine kinase family that mediates contact-dependent signaling between adjacent cells. EphA7 participates in Eph/ephrin bidirectional signaling to regulate cytoskeletal remodeling, cell adhesion, and directional migration, with established roles in tissue patterning and neural development. Downstream pathway outputs commonly intersect with Rho family GTPase signaling and MAPK/ERK and PI3K/AKT-associated programs that influence proliferation and survival. Altered EPHA7 expression or signaling has been reported across multiple cancer types and neurodevelopmental contexts, making it relevant for mechanistic studies of invasion, differentiation, and cell–cell communication.

EphA7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EPHA7 expression without altering the underlying DNA sequence.

EphA7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EPHA7 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 EPHA7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EphA7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EPHA7 locus and enabling the study of EphA7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EphA7 pathway restoration in tumor cells with silenced or reduced EPHA7 expression.

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