ART1 Knockout HCT 116 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population in which the ART1 gene has been disrupted via non-homologous end joining (NHEJ)-mediated insertions or deletions. This heterogeneous pool of HCT 116 cells carries diverse loss-of-function mutations across the ART1 locus, enabling robust functional studies without clonal selection artifacts. The polyclonal format preserves biological variability while effectively abolishing ART1 protein expression, creating a versatile model for investigating ADP-ribosyltransferase-dependent processes in a colorectal carcinoma background.
The HCT 116 cell line is an extensively characterized human colorectal adenocarcinoma model derived from a Dukes’ type C tumor. These epithelial cells retain key oncogenic mutations (e.g., KRAS G13D) and exhibit microsatellite instability (MSI), reflecting a mismatch repair-deficient phenotype common to a subset of colorectal cancers. HCT 116 cells are widely employed in cancer biology research due to their reproducible growth kinetics, metastatic potential in xenograft models, and responsiveness to immune and cytokine stimuli, making them particularly suited for studying the interplay between tumor-intrinsic ADP-ribosylation and the tumor microenvironment.
ART1 encodes a glycosylphosphatidylinositol (GPI)-anchored ecto-enzyme that catalyzes mono-ADP-ribosylation of arginine residues on extracellular and membrane-associated proteins. This post-translational modification is activated by upstream signals such as IFN-??, STAT1, and NF-??B, and it targets downstream substrates including integrin family members, cell adhesion molecules, and cytokine receptors. ART1 utilizes NAD+ as a co-substrate, and its activity modulates protein?Cprotein interactions and receptor function, thereby influencing immune cell adhesion, signal transduction, and apoptosis. Representative pathway components encompass ART1, NAD+, ADP-ribosylated target proteins, and immune signaling effectors, positioning ART1 at the intersection of NAD+ metabolism and immune regulatory networks.
In HCT 116 cells, ART1 knockout disrupts the arginine-specific ADP-ribosylation machinery, providing a clean loss-of-function model to dissect the enzyme’s contributions to colorectal carcinoma biology. Because HCT 116 cells express surface molecules that can be modified by ART1, ablation of ART1 may alter adhesive properties, immune recognition, or responses to inflammatory cytokines. This knockout model is especially relevant for examining how tumor cell-intrinsic ADP-ribosylation influences interactions with immune cells, matrix components, and soluble factors within the tumor microenvironment, offering insights into mechanisms of immune evasion and metastatic progression.
Researchers can employ ART1 Knockout HCT 116 Polyclonal Cells in a variety of experimental contexts, including western blotting and RT-qPCR for confirmation of gene disruption, ADP-ribosylation assays to quantify enzymatic activity loss, cell viability and apoptosis assays under immune challenge, migration assays to assess metastatic behavior, and RNA-seq for transcriptomic profiling. This model supports drug target validation studies, functional characterization of ADP-ribosylation in immune modulation, and exploration of tumor microenvironment crosstalk. For further details or customized knockout solutions, please contact Ascent Research.
