The ART1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This model provides a targeted disruption of the ART1 gene, ablating its function to study mono-ADP-ribosylation-mediated processes. The polyclonal pool format ensures a heterogeneous knockout background, minimizing clonal selection bias and enhancing reproducibility in downstream assays. These cells are particularly suited for examining ART1-dependent signaling and cell surface modification in intestinal epithelial cancer biology.
The parental HT29 line, established from a primary colorectal tumor of a 44-year-old female, serves as an intestinal epithelial model capable of differentiation and mucus secretion. HT29 cells retain critical colorectal epithelial characteristics, making them ideal for investigating tumor cell biology, host-microbe interactions, and mucosal immune responses. The ART1 knockout in this background thus allows researchers to explore the intersection of ADP-ribosylation and epithelial function in a clinically relevant setting.
ART1 is a GPI-anchored enzyme that catalyzes the transfer of ADP-ribose from NAD+ to arginine residues on target proteins, a modification that regulates immune cell adhesion and signaling. Its activity is potently induced by interferon-gamma (IFN-??) through the JAK/STAT pathway, downstream of the IFN-?? receptor. The integrin LFA-1 (CD11a/CD18) is a key downstream substrate; ADP-ribosylation of LFA-1 modulates its adhesive functions and immune synapse stability. ART1 also interacts with poly(ADP-ribose) glycohydrolase (PARG) and operates within lipid raft microdomains, suggesting spatial and dynamic control of this pathway. Thus, ART1 integrates signals from IFN-??, LFA-1, PARG, and lipid rafts to fine-tune immune cell interactions.
In HT29 colorectal adenocarcinoma cells, ART1 knockout eliminates cell-surface mono-ADP-ribosyltransferase activity, disrupting post-translational modifications critical for tumor-immune microenvironment crosstalk. Loss of ART1 is predicted to impair adhesion to immune cells, alter cytokine-driven signaling, and affect tumor cell migration, offering a model to dissect mechanisms of immune evasion in colorectal cancer. This system is also relevant for studying graft-versus-host disease and autoimmunity, where ADP-ribosylation of cell surface proteins plays a pathogenic role.
The ART1 Knockout HT29 Polyclonal Cells support a wide range of research applications, including western blotting and RT-qPCR for knockout validation, flow cytometry for surface marker and ADP-ribosylation profiling, and migration/invasion assays to evaluate metastatic potential. Co-culture with T cells or NK cells enables functional studies of immune synapse formation and evasion. Specialized assays can detect mono-ADP-ribosylated substrates. These tools are valuable for immunomodulatory drug screening and studying ADP-ribosylation biology in colorectal cancer. For technical support or ordering, contact Ascent Research.
