Fibulin-1 CRISPR Activation Plasmid (h): sc-401754-ACT

FBLN1 encodes fibulin-1, a secreted extracellular matrix glycoprotein that associates with elastic fibers and basement membrane components to regulate matrix assembly, cell adhesion, and tissue remodeling. Fibulin-1 interacts with integrins and other ECM proteins to influence cytoskeletal organization, cell migration, and growth factor availability, linking it to extracellular matrix–receptor signaling and mechanotransduction. Altered FBLN1 expression and fibulin-1 deposition have been observed in contexts of vascular remodeling, fibrosis, and tumor-associated stromal changes, making it relevant for studies of microenvironment-driven phenotypes. In human cell models, FBLN1 is frequently examined for its contributions to epithelial–mesenchymal interactions, endothelial function, and matrix-dependent regulation of proliferation and motility.

Fibulin-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FBLN1 expression without altering the underlying DNA sequence.

Fibulin-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FBLN1 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 FBLN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Fibulin-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FBLN1 locus and enabling the study of Fibulin-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Fibulin-1 pathway restoration in tumor cells with silenced or reduced FBLN1 expression.

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