SEMA3C CRISPR Activation Plasmid (h): sc-402489-ACT

SEMA3C encodes Semaphorin-3C, a secreted guidance cue that signals through neuropilin and plexin receptors to regulate cytoskeletal dynamics, directional migration, and cell–cell interactions. Beyond axon guidance, SEMA3C contributes to vascular patterning and tissue remodeling by modulating pathways that intersect with Rho-family GTPases, focal adhesion turnover, and integrin-dependent adhesion. Dysregulated SEMA3C expression has been reported in contexts of aberrant angiogenic signaling, invasive cell behavior, and altered stromal–tumor communication, making it relevant for mechanistic studies of developmental and disease-associated microenvironments. Its activity is commonly examined in models of epithelial and endothelial plasticity, extracellular matrix organization, and receptor-mediated semaphorin signaling.

SEMA3C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SEMA3C expression without altering the underlying DNA sequence.

SEMA3C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SEMA3C 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 SEMA3C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SEMA3C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SEMA3C locus and enabling the study of SEMA3C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SEMA3C pathway restoration in tumor cells with silenced or reduced SEMA3C expression.

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