elastin CRISPR Activation Plasmid (h): sc-400640-ACT

Human ELN encodes elastin, a highly crosslinked extracellular matrix protein that provides elasticity and recoil to tissues such as arterial walls, lung, and skin. Elastin is deposited as tropoelastin and assembled onto microfibrillar scaffolds, coupling matrix organization with smooth muscle cell behavior, mechanotransduction, and extracellular matrix remodeling pathways. ELN expression and elastin fiber integrity influence vascular compliance and connective tissue homeostasis, linking altered elastogenesis to heritable and acquired disorders characterized by arterial stenosis, aneurysm susceptibility, and dermal laxity. As a matrix component with long half-life, elastin is frequently studied in models of development, aging, and fibrosis-associated changes in tissue biomechanics.

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

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

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