SCUBE2 CRISPR Activation Plasmid (h): sc-410079-ACT

SCUBE2 (signal peptide, CUB domain, EGF-like domain-containing 2) encodes a secreted cell-surface–associated glycoprotein that modulates extracellular signaling and cell–cell communication. Through its CUB and EGF-like domains, SCUBE2 is implicated in regulating growth factor availability and receptor signaling dynamics, with reported connections to developmental programs and endothelial or epithelial cell behaviors. Altered SCUBE2 expression has been associated with dysregulated proliferation, migration, and differentiation in multiple disease-relevant contexts, including cancer biology and vascular remodeling. As a pathway-linked extracellular regulator, SCUBE2 is frequently studied for its impact on signaling networks that shape tissue organization and tumor microenvironment interactions.

SCUBE2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SCUBE2 expression without altering the underlying DNA sequence.

SCUBE2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SCUBE2 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 SCUBE2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SCUBE2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SCUBE2 locus and enabling the study of SCUBE2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SCUBE2 pathway restoration in tumor cells with silenced or reduced SCUBE2 expression.

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