TACE/ADAM17 CRISPR Activation Plasmid (h): sc-400827-ACT

ADAM17, also known as TACE, is a membrane-anchored metalloprotease that catalyzes ectodomain shedding of diverse substrates including TNF-α, EGFR ligands (such as amphiregulin and TGF-α), and multiple adhesion molecules. Through regulated proteolysis at the cell surface, TACE/ADAM17 integrates inflammatory signaling with growth factor–dependent pathways including EGFR–MAPK/ERK and NF-κB, shaping cytokine release, receptor activation, and tissue remodeling. Dysregulated ADAM17 activity has been linked to chronic inflammation and cancer-related signaling networks, and is widely studied for its role in immune cell communication, epithelial homeostasis, and microenvironmental crosstalk.

TACE/ADAM17 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAM17 expression without altering the underlying DNA sequence.

TACE/ADAM17 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAM17 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 ADAM17 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TACE/ADAM17 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAM17 locus and enabling the study of TACE/ADAM17-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TACE/ADAM17 pathway restoration in tumor cells with silenced or reduced ADAM17 expression.

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