SAV1 CRISPR Activation Plasmid (h): sc-402600-ACT

Human SAV1 (Salvador family WW domain containing protein 1) is a core scaffold of the Hippo tumor suppressor pathway that coordinates MST1/2 and LATS1/2 kinase signaling to restrain YAP/TAZ-driven transcriptional programs. By promoting phosphorylation-dependent cytoplasmic retention and turnover of YAP/TAZ, SAV1 helps regulate contact inhibition, proliferation control, apoptosis, and tissue homeostasis. Altered SAV1 expression or Hippo pathway dysfunction has been linked to dysregulated cell growth and differentiation states in multiple disease-relevant contexts, particularly those characterized by aberrant YAP/TAZ activity. SAV1 is therefore widely studied in pathways intersecting with cytoskeletal dynamics, cell polarity, mechanotransduction, and organ size control.

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

SAV1 CRISPR Activation Plasmid (h) 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.