Septin 4 CRISPR Activation Plasmid (h): sc-402288-ACT

Human SEPT4 encodes septin 4, a GTP-binding cytoskeletal scaffolding protein that assembles into hetero-oligomeric filaments and ring-like structures to organize membrane domains. Septin 4 contributes to cytokinesis, midbody maturation, vesicle trafficking, and establishment of diffusion barriers by coordinating with actin, microtubules, and associated regulators of cell polarity. Through these roles, SEPT4 helps control cell division fidelity and cytoskeletal remodeling programs that influence neuronal and epithelial homeostasis. Dysregulated septin organization or SEPT4 expression has been linked in the literature to neurodegenerative processes, altered apoptosis signaling, and cancer-associated changes in proliferation and migration, supporting its use as a mechanistic node in cytoskeletal and cell-cycle research.

Septin 4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SEPT4 expression without altering the underlying DNA sequence.

Septin 4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SEPT4 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 SEPT4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Septin 4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SEPT4 locus and enabling the study of Septin 4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Septin 4 pathway restoration in tumor cells with silenced or reduced SEPT4 expression.

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