Fibulin-5 CRISPR Activation Plasmid (h): sc-402597-ACT

FBLN5 encodes fibulin-5, an extracellular matrix glycoprotein that binds elastin and integrins to support elastic fiber assembly and tissue elasticity in skin, vasculature, and lung. Fibulin-5 helps organize microfibrillar architecture and modulates cell–matrix adhesion, influencing processes such as smooth muscle cell behavior, angiogenesis, and matrix remodeling through pathways linked to ECM–receptor interactions and protease regulation. Altered FBLN5 expression or function has been associated with dysregulated elastogenesis, connective tissue fragility, and degenerative changes in elastic tissues, making it a relevant target for mechanistic studies of ECM homeostasis. In human model systems, FBLN5 perturbation is used to probe how extracellular matrix composition shapes cellular signaling, biomechanics, and tissue-level phenotypes.

Fibulin-5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FBLN5 expression without altering the underlying DNA sequence.

Fibulin-5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FBLN5 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 FBLN5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Fibulin-5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FBLN5 locus and enabling the study of Fibulin-5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Fibulin-5 pathway restoration in tumor cells with silenced or reduced FBLN5 expression.

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