Fibromodulin CRISPR Activation Plasmid (h): sc-401610-ACT

FMOD encodes fibromodulin, a secreted small leucine-rich proteoglycan enriched in connective tissues where it binds fibrillar collagens and regulates extracellular matrix organization. Fibromodulin contributes to collagen fibrillogenesis, matrix assembly, and mechanobiology, influencing cell adhesion, migration, and tissue remodeling programs. Through its interactions with matrix components and growth factor signaling such as TGF-β–linked pathways, FMOD can modulate fibroblast behavior and stromal responses. Altered FMOD expression and matrix remodeling are associated with fibrotic phenotypes and tumor microenvironment dynamics, making it a useful target for studying ECM-driven disease mechanisms.

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

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

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