FAM20C CRISPR Activation Plasmid (h): sc-406736-ACT

FAM20C encodes a secreted kinase localized to the Golgi apparatus that phosphorylates Ser-x-Glu/pSer motifs on numerous extracellular and secreted proteins, including members of the SIBLING family and other matrix-associated factors. Through regulation of protein phosphorylation in the secretory pathway, FAM20C influences extracellular matrix organization, biomineralization, and cell–matrix signaling with downstream effects on osteogenic differentiation and tissue homeostasis. Altered FAM20C activity and dysregulated secreted phosphoproteomes have been linked to defects in skeletal and dental development and broader connective tissue phenotypes, making it a key node for studying matrix biology. Its role in Golgi-dependent post-translational modification also connects FAM20C to pathways controlling secretion, protein quality control, and intercellular signaling via the extracellular milieu.

FAM20C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FAM20C expression without altering the underlying DNA sequence.

FAM20C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FAM20C 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 FAM20C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FAM20C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FAM20C locus and enabling the study of FAM20C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FAM20C pathway restoration in tumor cells with silenced or reduced FAM20C expression.

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