Sclerostin CRISPR Activation Plasmid (m): sc-428914-ACT

Mouse Sost encodes sclerostin, a secreted glycoprotein predominantly produced by osteocytes that functions as a potent inhibitor of canonical Wnt/β-catenin signaling in bone. By binding the LRP5/6 co-receptors, sclerostin antagonizes Wnt ligand–mediated receptor activation, constraining osteoblast differentiation, matrix deposition, and skeletal mineralization while contributing to mechanotransduction-dependent remodeling. Sost also interfaces with BMP and TGF-β signaling crosstalk that shapes osteogenic gene programs and bone homeostasis. Dysregulated sclerostin activity is implicated in altered bone mass phenotypes and provides a mechanistic entry point for modeling skeletal development, remodeling, and osteoimmunology-associated changes in bone turnover.

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

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

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