LOXL2 CRISPR Activation Plasmid (h): sc-401021-ACT

Human LOXL2 (lysyl oxidase like 2) is a secreted copper-dependent amine oxidase that catalyzes oxidative deamination of lysine residues in collagen and elastin, promoting covalent crosslinking and extracellular matrix (ECM) maturation. Through ECM remodeling, LOXL2 regulates cell adhesion, migration, and mechanotransduction, interfacing with pathways linked to epithelial–mesenchymal transition, tissue fibrosis, and stromal remodeling. Dysregulated LOXL2 expression and activity are frequently associated with altered matrix stiffness and invasive phenotypes in cancer biology, as well as fibrotic and inflammatory microenvironments. These functions make LOXL2 a useful target for studying ECM-driven regulation of cell behavior, metastasis-related processes, and microenvironmental signaling.

LOXL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LOXL2 expression without altering the underlying DNA sequence.

LOXL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LOXL2 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 LOXL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LOXL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LOXL2 locus and enabling the study of LOXL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LOXL2 pathway restoration in tumor cells with silenced or reduced LOXL2 expression.

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