SEMA3F CRISPR Activation Plasmid (h): sc-403710-ACT

SEMA3F encodes semaphorin-3F, a secreted guidance cue that signals primarily through neuropilin and plexin receptors to regulate cytoskeletal remodeling, directional cell migration, and axon pathfinding. In non-neuronal tissues, SEMA3F participates in pathways controlling cell–cell communication, adhesion dynamics, and vascular patterning, linking it to processes such as tissue organization and microenvironmental signaling. Altered SEMA3F expression has been associated with dysregulated motility and invasive behavior in cancer biology, and it is also studied in neurodevelopmental and neuroinflammatory contexts where guidance cues shape circuit formation and immune–neuronal cross-talk. As a ligand-driven regulator of semaphorin signaling, SEMA3F provides a tractable node for probing receptor-mediated guidance and migration programs.

SEMA3F CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SEMA3F expression without altering the underlying DNA sequence.

SEMA3F CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SEMA3F 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 SEMA3F transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SEMA3F expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SEMA3F locus and enabling the study of SEMA3F-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SEMA3F pathway restoration in tumor cells with silenced or reduced SEMA3F expression.

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