The ATOX1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ATOX1 gene in the HT29 human colorectal adenocarcinoma cell line. This heterogeneous pool of cells, generated through CRISPR/Cas9-mediated gene disruption, provides a loss-of-function model for investigating ATOX1-dependent processes. Unlike clonal isolates, the polyclonal format captures a spectrum of genetic modifications, facilitating robust functional studies while minimizing clonal artifacts. The product is suited for advanced research in copper biology, oxidative stress, and cancer cell signaling.
HT29 cells are derived from a primary grade II colorectal adenocarcinoma from a 44-year-old female. As a widely used intestinal epithelial model, HT29 cells retain the capacity to differentiate into enterocyte-like cells, making them valuable for studying colorectal cancer initiation, progression, and drug response. Their epithelial origin supports investigations of cell polarity, barrier function, and tumor-stroma interactions. The knockout in this background enables dissection of copper-related pathways specifically within the context of colorectal adenocarcinoma, a disease increasingly linked to dysregulated copper metabolism.
ATOX1 encodes a copper chaperone that facilitates the delivery of copper ions to the P-type ATPases ATP7A and ATP7B, which are essential for copper secretion and intracellular distribution. ATOX1 also supplies copper to cuproenzymes such as superoxide dismutase 3 (SOD3), a key extracellular antioxidant. Copper homeostasis involves a network of proteins including the copper importer CTR1 and the CCS chaperone for SOD1. Upstream, ATOX1 expression is regulated by intracellular copper levels, oxidative stress, and the transcription factor HIF1A. Downstream targets include ATP7A, ATP7B, and copper-dependent enzymes. ATOX1 interacts directly with ATP7A, ATP7B, and copper ions. Loss of ATOX1 function abolishes copper delivery to these transporters, disrupting copper homeostasis, reducing antioxidant capacity, and promoting oxidative stress, which may alter cell proliferation and migration, potentially via PI3K/AKT signaling.
In the HT29 colorectal cancer model, ATOX1 knockout enables precise investigation of copper trafficking and its impact on tumor biology. Colorectal adenocarcinomas often exhibit altered copper levels and oxidative stress responses; ATOX1 disruption can reveal how copper dysregulation modulates redox balance, cell survival, and metastatic potential. The knockout cells are particularly relevant for studying the interplay between copper metabolism and drug resistance, including resistance to platinum-based chemotherapeutics that rely on copper-related pathways. Moreover, the model may illuminate the role of copper in shaping the tumor microenvironment, where secreted cuproenzymes and copper-dependent signaling contribute to cancer progression.
This polyclonal knockout population supports diverse experimental workflows. Researchers can validate ATOX1 disruption via western blotting for ATOX1 and its targets ATP7A/ATP7B, RT-qPCR, and copper uptake assays. Functional readouts include ROS detection using DCFDA, cell viability assays (MTT/CCK-8), and migration/invasion assays to assess metastatic behavior. Immunofluorescence can trace copper trafficking, and phospho-AKT/ERK analysis can probe associated signaling changes. Typical applications encompass copper homeostasis studies in colorectal cancer, oxidative stress modulation, tumor microenvironment interactions, and drug resistance mechanisms. For technical inquiries or ordering, contact Ascent Research.
