LOXL4 CRISPR Activation Plasmid (h): sc-403384-ACT

Human LOXL4 (lysyl oxidase like 4) is a copper-dependent amine oxidase that catalyzes oxidative deamination of lysine residues in collagens and elastin, promoting covalent crosslinking and maturation of the extracellular matrix. Through matrix remodeling, LOXL4 influences tissue stiffness, cell adhesion, and migration, integrating with ECM–integrin signaling and broader wound repair and fibrotic processes. Altered LOXL4 expression has been reported across multiple solid tumors and in remodeling-associated pathologies, where changes in collagen architecture can affect invasion, angiogenesis, and stromal-epithelial interactions. These features make LOXL4 a useful node for studying extracellular matrix organization and microenvironment-dependent phenotypes in human cell models.

LOXL4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LOXL4 expression without altering the underlying DNA sequence.

LOXL4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LOXL4 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 LOXL4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LOXL4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LOXL4 locus and enabling the study of LOXL4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LOXL4 pathway restoration in tumor cells with silenced or reduced LOXL4 expression.

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