SEMA3E CRISPR Activation Plasmid (h): sc-404797-ACT

SEMA3E encodes semaphorin-3E, a secreted guidance cue that signals primarily through plexin receptors to regulate cytoskeletal remodeling, cell migration, and directional navigation. In addition to roles in axon guidance, SEMA3E influences vascular patterning and endothelial behavior, integrating with pathways that control adhesion dynamics and Rho family GTPase activity. Dysregulated SEMA3E signaling has been associated with altered invasive potential, angiogenic responses, and remodeling of the tissue microenvironment in multiple disease contexts, making it a relevant node for studying motility and developmental signaling. These functions position SEMA3E as a useful molecular handle for probing how extracellular guidance cues shape cell-state transitions and tissue organization.

SEMA3E CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SEMA3E expression without altering the underlying DNA sequence.

SEMA3E CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SEMA3E 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 SEMA3E transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SEMA3E expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SEMA3E locus and enabling the study of SEMA3E-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SEMA3E pathway restoration in tumor cells with silenced or reduced SEMA3E expression.

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