Sc1 CRISPR Activation Plasmid (m): sc-420099-ACT

Mouse Sparcl1 encodes Sc1 (SPARC-like 1), a secreted matricellular glycoprotein that modulates extracellular matrix organization and cell–matrix signaling. Sc1 influences cell adhesion, migration, and proliferation by shaping interactions between stromal cells and the surrounding matrix, with downstream effects on tissue remodeling and vascular-associated processes. Sparcl1 expression is frequently linked to changes in stromal architecture and microenvironmental cues that intersect with wound repair and fibrosis-like remodeling programs. In biomedical research, Sparcl1/Sc1 is used to study how extracellular matrix composition and matricellular signaling coordinate tissue homeostasis and dysregulated remodeling observed across diverse disease models.

Sc1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Sparcl1 expression without altering the underlying DNA sequence.

Sc1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Sparcl1 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 Sparcl1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sc1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Sparcl1 locus and enabling the study of Sc1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sc1 pathway restoration in tumor cells with silenced or reduced Sparcl1 expression.

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