SPARCL1 CRISPR Activation Plasmid (h): sc-404048-ACT

SPARCL1 (SPARC-like protein 1, also known as hevin) is a secreted matricellular glycoprotein that modulates cell–matrix interactions and extracellular matrix (ECM) organization in human tissues. It influences processes such as cell adhesion, migration, and tissue remodeling through interactions with structural ECM components and regulation of signaling in the perivascular and stromal microenvironment. SPARCL1 expression is frequently altered across contexts involving vascular biology and tumor-associated stroma, where ECM composition and mechanochemical cues shape cellular behavior. As a readout and driver of microenvironmental state, SPARCL1 is widely studied in models of angiogenesis, inflammation-associated remodeling, and cancer biology.

SPARCL1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SPARCL1 expression without altering the underlying DNA sequence.

SPARCL1 CRISPR Activation Plasmid (h) 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 SPARCL1 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 SPARCL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SPARCL1 pathway restoration in tumor cells with silenced or reduced SPARCL1 expression.

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