LOXL1 CRISPR Activation Plasmid (h): sc-401965-ACT

LOXL1 encodes lysyl oxidase–like 1, a secreted copper-dependent amine oxidase that catalyzes oxidative deamination of lysine residues in collagen and elastin, enabling covalent crosslinking and stabilization of extracellular matrix architecture. By regulating elastic fiber assembly and matrix remodeling, LOXL1 influences tissue mechanics, cell–matrix signaling, and processes such as fibrosis and wound repair. Altered LOXL1 activity has been associated with connective tissue and ocular extracellular matrix disorders, including exfoliation syndrome/glaucoma risk, and is studied in the context of vascular and pelvic floor tissue integrity. As a matrix-modifying enzyme, LOXL1 serves as a useful node for interrogating ECM homeostasis, mechanobiology, and remodeling-driven disease phenotypes in human cell models.

LOXL1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LOXL1 expression without altering the underlying DNA sequence.

LOXL1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LOXL1 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 LOXL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LOXL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LOXL1 locus and enabling the study of LOXL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LOXL1 pathway restoration in tumor cells with silenced or reduced LOXL1 expression.

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