EMILIN-1 CRISPR Activation Plasmid (h): sc-403434-ACT

EMILIN1 encodes EMILIN-1, an extracellular matrix glycoprotein enriched in elastic fiber–associated microfibrils that contributes to tissue architecture and biomechanical signaling in the vasculature and connective tissues. EMILIN-1 modulates cell–matrix interactions and influences processes such as cell adhesion, migration, and vascular homeostasis, with reported links to regulation of growth factor signaling in the extracellular milieu. Altered EMILIN1 expression or extracellular matrix organization is relevant to studies of vascular remodeling, fibrosis, and tumor–stroma interactions where matrix composition can shape cellular phenotype. As a matricellular component, EMILIN-1 is frequently examined in pathways governing extracellular matrix assembly and mechanotransduction.

EMILIN-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EMILIN1 expression without altering the underlying DNA sequence.

EMILIN-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EMILIN1 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 EMILIN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EMILIN-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EMILIN1 locus and enabling the study of EMILIN-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EMILIN-1 pathway restoration in tumor cells with silenced or reduced EMILIN1 expression.

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