SEMA3D CRISPR Activation Plasmid (h): sc-403924-ACT

SEMA3D encodes semaphorin 3D, a secreted guidance cue that signals through neuropilin and plexin receptor complexes to regulate cytoskeletal remodeling, directional cell migration, and axon pathfinding. In human tissues, SEMA3D contributes to tissue patterning and vascular and neural network organization by modulating Rho family GTPase activity and integrin-dependent adhesion. Dysregulated semaphorin signaling is associated with altered cell motility and microenvironmental interactions observed across developmental disorders and cancer-related processes, making SEMA3D a useful node for studying invasion, angiogenesis, and neural wiring. Researchers commonly interrogate SEMA3D function in pathways governing guidance signaling, epithelial–mesenchymal dynamics, and context-dependent regulation of growth factor and extracellular matrix responses.

SEMA3D CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SEMA3D expression without altering the underlying DNA sequence.

SEMA3D CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SEMA3D 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 SEMA3D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SEMA3D expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SEMA3D locus and enabling the study of SEMA3D-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SEMA3D pathway restoration in tumor cells with silenced or reduced SEMA3D expression.

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