Testican-1 CRISPR Activation Plasmid (h): sc-406915-ACT

SPOCK1 encodes the human extracellular matrix proteoglycan Testican-1, a secreted and pericellular glycoprotein that modulates matrix organization and cell–matrix interactions. Testican-1 has been implicated in regulation of protease activity and remodeling processes that influence cell adhesion, migration, neurite outgrowth, and tissue architecture, linking it to pathways governing extracellular matrix dynamics and cellular motility programs. Altered SPOCK1 expression has been reported in multiple disease contexts, including cancers and fibrotic or inflammatory states, where dysregulated matrix remodeling and invasive behavior are key biological features. As a result, SPOCK1/Testican-1 is frequently studied as a context-dependent regulator of microenvironmental signaling and phenotypic plasticity.

Testican-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SPOCK1 expression without altering the underlying DNA sequence.

Testican-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SPOCK1 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 SPOCK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Testican-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SPOCK1 locus and enabling the study of Testican-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Testican-1 pathway restoration in tumor cells with silenced or reduced SPOCK1 expression.

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