TEM8 CRISPR Activation Plasmid (h2): sc-404903-ACT-2

Human ANTXR1 encodes tumor endothelial marker 8 (TEM8), a cell-surface anthrax toxin receptor that functions as an adhesion- and matrix-interacting protein implicated in endothelial cell behavior and tissue remodeling. TEM8 binds extracellular matrix components and contributes to cytoskeletal organization and receptor-mediated endocytosis, linking ANTXR1 activity to angiogenic programs and microenvironmental signaling. Genetic and functional studies associate ANTXR1 with vascular development phenotypes and extracellular matrix dysregulation, with relevance to tumor-associated vasculature and disorders affecting connective tissue homeostasis. Gene editing of ANTXR1 enables mechanistic interrogation of TEM8-dependent trafficking and adhesion pathways, analysis of endothelial–matrix interactions, and development of isogenic cellular models for pathway and functional genomics studies.

TEM8 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous ANTXR1 expression without altering the underlying DNA sequence.

TEM8 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ANTXR1 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 ANTXR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TEM8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ANTXR1 locus and enabling the study of TEM8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TEM8 pathway restoration in tumor cells with silenced or reduced ANTXR1 expression.

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