SAV1 CRISPR Activation Plasmid (m): sc-425662-ACT

Sav1 encodes Salvador family WW domain-containing protein 1 (SAV1), a scaffold component of the Hippo signaling pathway that cooperates with MST1/2 and LATS kinases to regulate YAP/TAZ activity and transcriptional outputs controlling proliferation, apoptosis, and tissue growth. In mouse cells, SAV1 supports contact inhibition and epithelial homeostasis by coordinating kinase complex assembly and phosphorylation cascades that restrain pro-growth gene expression programs. Dysregulation of SAV1-linked Hippo signaling is associated with altered organ size control, aberrant regenerative responses, and tumor-associated phenotypes in experimental models, making Sav1 a useful node for studying growth control and mechanotransduction. SAV1 also connects to cytoskeletal dynamics and junctional signaling, providing a route to interrogate how cell polarity and mechanical cues influence transcriptional state.

SAV1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Sav1 expression without altering the underlying DNA sequence.

SAV1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Sav1 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 Sav1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SAV1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Sav1 locus and enabling the study of SAV1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SAV1 pathway restoration in tumor cells with silenced or reduced Sav1 expression.

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