EphA1 CRISPR Activation Plasmid (h): sc-403340-ACT

EPHA1 encodes EphA1, a receptor tyrosine kinase in the ephrin-A signaling network that governs contact-dependent cell communication, cytoskeletal remodeling, and directional cell migration. Upon ephrin ligand engagement, EphA1 triggers bidirectional signaling that intersects with Rho-family GTPases, MAPK/ERK, and PI3K/AKT pathways to shape adhesion, boundary formation, and epithelial tissue organization. Dysregulated EPHA1 expression or signaling has been associated with altered cell–cell interactions and invasive phenotypes in multiple disease contexts, supporting its use as a mechanistic node for studying microenvironment-driven signaling. EphA1 is also implicated in processes linked to angiogenesis and immune–stromal crosstalk, making it relevant for pathway-focused functional genomics in human cells.

EphA1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EPHA1 expression without altering the underlying DNA sequence.

EphA1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EPHA1 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 EPHA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EphA1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EPHA1 locus and enabling the study of EphA1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EphA1 pathway restoration in tumor cells with silenced or reduced EPHA1 expression.

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