ADAMTS-L2 CRISPR Activation Plasmid (h): sc-411325-ACT

ADAMTSL2 encodes ADAMTS-L2, a secreted extracellular matrix glycoprotein in the ADAMTS-like family that lacks catalytic protease activity but modulates matrix organization and cell–matrix signaling. ADAMTS-L2 participates in extracellular microfibril biology and is functionally linked to transforming growth factor-β (TGF-β) regulation through interactions with fibrillin-rich matrices, influencing tissue architecture and mechanical homeostasis. Altered ADAMTSL2 expression or function is associated with connective tissue phenotypes and dysregulated matrix remodeling, making it relevant for studies of fibrosis-like programs, developmental morphogenesis, and stromal cell biology. Its extracellular localization also supports investigation of paracrine signaling, matrix deposition, and remodeling dynamics in human cell and organoid systems.

ADAMTS-L2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAMTSL2 expression without altering the underlying DNA sequence.

ADAMTS-L2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAMTSL2 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 ADAMTSL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAMTS-L2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAMTSL2 locus and enabling the study of ADAMTS-L2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAMTS-L2 pathway restoration in tumor cells with silenced or reduced ADAMTSL2 expression.

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