Fibulin-4 CRISPR Activation Plasmid (h): sc-404037-ACT

EFEMP2 encodes fibulin-4, a secreted extracellular matrix glycoprotein that integrates into elastic fiber microfibrils and supports elastogenesis through interactions with tropoelastin, fibrillins, and lysyl oxidase–dependent crosslinking machinery. Fibulin-4 contributes to extracellular matrix organization, mechanotransduction, and vascular wall integrity, influencing connective tissue remodeling and smooth muscle cell homeostasis. Altered EFEMP2 expression or fibulin-4 dysfunction is linked to disrupted elastic fiber assembly and extracellular matrix defects associated with heritable connective tissue and vascular phenotypes. These biology- and matrix-centric roles make EFEMP2 a useful node for studying elastic fiber pathways, ECM proteostasis, and cell–matrix signaling in human model systems.

Fibulin-4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EFEMP2 expression without altering the underlying DNA sequence.

Fibulin-4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EFEMP2 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 EFEMP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Fibulin-4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EFEMP2 locus and enabling the study of Fibulin-4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Fibulin-4 pathway restoration in tumor cells with silenced or reduced EFEMP2 expression.

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