This product is a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human ART1 gene in the NCI-H1299 non-small cell lung cancer (NSCLC) line. The ART1 gene has been disrupted through CRISPR/Cas9-mediated gene editing, resulting in a heterogeneous cell pool carrying loss-of-function mutations. This polyclonal knockout model enables robust investigation of ART1-catalyzed ADP-ribosylation in cancer biology, free from clonal selection bias.
The NCI-H1299 cell line is a human lung adenocarcinoma epithelial line derived from a lymph node metastasis. It is characterized by a homozygous TP53 deletion (p53 null) and a KRAS G12C oncogenic mutation, conferring an aggressive metastatic phenotype. NCI-H1299 is extensively used to study tumor invasion, metastasis, and therapeutic resistance, making it a relevant host for examining ART1’s role in metastatic progression.
ART1 encodes an arginine-specific ADP-ribosyltransferase that utilizes NAD+ to modify cell surface proteins such as the purinergic receptor P2X7R and integrin ??1. This post-translational modification regulates apoptosis, cell adhesion, and immune signaling. ART1 expression is activated by IFN-??, NF-??B, and STAT3, and it functions in pathways linking NAD+ metabolism, purinergic signaling, and integrin-mediated adhesion. Downstream, integrin ??1 ADP-ribosylation impacts FAK phosphorylation and focal adhesion dynamics, while P2X7R modification influences calcium flux and T cell apoptosis. ART1 operates within lipid raft domains, forming complexes with interacting partners that modulate its substrate specificity.
In the context of NCI-H1299 cells, ART1 knockout is expected to disrupt surface ADP-ribosylation of P2X7R and integrins, leading to altered apoptotic responses, impaired adhesion to extracellular matrix components, and reduced migratory capacity. This model provides a powerful system to investigate how ART1-dependent signaling contributes to lung adenocarcinoma metastasis, especially in cells with p53 deficiency and oncogenic KRAS. The combination of ART1 loss with these genetic alterations may reveal synergistic pathways driving tumor aggressiveness.
Typical applications include mechanistic studies of ADP-ribosylation within the tumor microenvironment, investigation of purinergic signaling in cancer progression, analysis of cell adhesion and migration pathways, and drug sensitivity profiling targeting NAD+ metabolism or ADP-ribosyltransferase activity. Researchers commonly employ western blotting for ART1 and ADP-ribosylation targets, RT-qPCR, immunofluorescence for ART1 localization, flow cytometry for integrins and apoptosis, Transwell migration/invasion assays, adhesion assays, phospho-FAK analysis, drug sensitivity assays with NAD+ metabolism inhibitors, RNA-seq, and Annexin V staining. For further information or custom inquiries, please contact Ascent Research.
