The ATG2A Knockout HT29 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, engineered to disrupt the ATG2A gene. This model provides a loss-of-function system for studying the lipid transfer protein ATG2A, which is essential for autophagosome biogenesis. The polyclonal format offers heterogeneity in gene-editing outcomes across the population, enabling robust, population-level analyses of autophagic function without the clonal artifacts associated with single-cell-derived lines. Researchers can utilize these cells to investigate the consequences of ATG2A disruption on autophagy and associated cellular processes in a well-characterized colorectal cancer background.
HT29 cells are an adherent, epithelial-like cell line originally established from a primary colorectal adenocarcinoma of a 44-year-old female. They retain key characteristics of the tissue of origin, including intact epithelial barrier properties and tumorigenic potential, making them a widely used model for colorectal cancer research. The cells form polarized monolayers and express markers typical of intestinal epithelium, providing a physiologically relevant context for studying autophagy. This host cell line is particularly suitable for examining the interplay between autophagy, cell polarity, and tumor cell biology, as it recapitulates many aspects of colorectal adenocarcinoma in vitro.
ATG2A functions as a lipid transfer protein at endoplasmic reticulum?Cphagophore contact sites, where it delivers phospholipids to the expanding autophagosomal membrane. Its activity is regulated upstream by the mTORC1-AMPK-ULK1 signaling axis in response to nutrient deprivation, and it interacts with WIPI4, ATG9A, and PI3P to facilitate membrane expansion. Downstream, ATG2A-mediated lipid transfer is required for the lipidation of LC3 and GABARAP family proteins, a critical step in autophagosome maturation and subsequent lysosomal degradation. The pathway includes the ULK1 complex, the PI3KC3 complex, WIPI2, ATG16L1, and the ATG5-ATG12 conjugation system, all converging on ATG2A-dependent autophagosome formation. Knockout of ATG2A thus disrupts autophagic flux, leading to accumulation of protein aggregates and damaged organelles.
In the context of HT29 colorectal adenocarcinoma cells, ATG2A knockout offers a powerful tool to dissect the role of autophagy in tumorigenesis, cell survival under metabolic stress, and resistance to chemotherapeutic agents. Colorectal cancer cells often exploit autophagy to cope with nutrient limitation within the tumor microenvironment and to evade apoptosis induced by drugs such as 5-fluorouracil (5-FU) and oxaliplatin. Impairment of ATG2A-dependent autophagosome biogenesis in these cells is expected to compromise their ability to degrade cytotoxic protein aggregates and recycle damaged mitochondria, leading to reduced viability under starvation and increased sensitivity to chemotherapy. This model therefore enables the investigation of autophagy as a therapeutic vulnerability in colorectal cancer.
Typical research applications include mechanistic studies of autophagosome biogenesis, screening of autophagy modulators, functional genomics analyses to identify genes that interact with ATG2A, and evaluation of ATG2A’s role in tumor progression and drug response. Researchers can employ a variety of assays, such as Western blotting for LC3-II and p62 accumulation, immunofluorescence staining of LC3 puncta and WIPI4 localization, autophagic flux assays using chloroquine, co-immunoprecipitation with WIPI4, and cell viability measurements under starvation conditions. Additional experiments may involve drug sensitivity testing with 5-FU or oxaliplatin, colony formation assays, and lipid transfer activity analyses. For additional technical specifications or to request a quotation, please contact Ascent Research.
