ART1 CRISPR Activation Plasmid (h): sc-403742-ACT

Human ART1 encodes ecto-ADP-ribosyltransferase 1, a GPI-anchored cell-surface enzyme that catalyzes mono-ADP-ribosylation of extracellular and membrane-associated proteins using NAD+ as a substrate. This post-translational modification can alter receptor function, adhesion, and immune-related signaling at the cell interface, linking ART1 to regulation of cell communication and stress responses. ART1 activity has been studied in the context of inflammatory microenvironments and cancer-associated phenotypes, where changes in cell-surface ADP-ribosylation may influence tumor–immune interactions and metastatic behavior. As a mediator of extracellular NAD+-dependent signaling, ART1 provides a tractable entry point for probing how ADP-ribosylation shapes pathway crosstalk in human cells.

ART1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ART1 expression without altering the underlying DNA sequence.

ART1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ART1 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 ART1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ART1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ART1 locus and enabling the study of ART1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ART1 pathway restoration in tumor cells with silenced or reduced ART1 expression.

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