SPARC CRISPR Activation Plasmid (m): sc-423101-ACT

Mouse Sparc encodes SPARC, a secreted matricellular glycoprotein that modulates extracellular matrix (ECM) organization, collagen deposition, and cell–matrix interactions. SPARC regulates adhesion, migration, and proliferation by shaping integrin signaling and growth factor availability, influencing processes such as wound repair, angiogenesis, and tissue remodeling. It is frequently linked to pathways governing fibrosis and stromal responses, and its dysregulation is associated with altered ECM homeostasis and inflammatory microenvironments. These properties make SPARC a useful node for studying mesenchymal cell behavior, ECM-driven signaling crosstalk, and microenvironmental regulation in disease-relevant models.

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

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

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