The ATG2B Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma epithelial cell line, engineered for targeted disruption of the ATG2B gene. This loss-of-function model is provided as a polyclonal cell population, enabling robust investigation of ATG2B-dependent processes without clonal selection artifacts. The knockout genotype is achieved through CRISPR/Cas9-mediated gene disruption, abolishing wild-type ATG2B protein expression while preserving the heterogeneous genomic integration profile characteristic of polyclonal editing. Researchers are advised to confirm knockout efficiency through appropriate genotyping and expression analyses prior to experimental use.
The HT29 host cell line is a widely employed model in colorectal cancer research, originating from a human colon adenocarcinoma with a well-characterized mutant p53 background. These adherent epithelial cells are extensively used to study colon cancer progression, intestinal epithelial barrier function, and cancer biology. Their genetic profile, including p53 mutation, makes them particularly relevant for investigating autophagy dysregulation in tumorigenesis, as impaired p53 signaling often coexists with altered autophagic activity. The HT29 line’s ability to form tight junctions and maintain epithelial polarity further supports its application in barrier integrity and metastasis studies.
ATG2B encodes a critical lipid transfer protein essential for autophagosome biogenesis, mediating the bulk transfer of phospholipids from the endoplasmic reticulum to the expanding phagophore membrane. The mechanistic summary highlights that ATG2B functions downstream of the ULK1-ATG13-RB1CC1 kinase complex and phosphatidylinositol-3-kinase (PIK3C3-ATG14) nucleation machinery, with recruitment to phagophore assembly sites mediated by WIPI family proteins (WIPI1, WIPI2, and WIPI4). Upstream regulators include the MTOR-AMPK axis and the ULK1 initiator complex, while ATG2B acts upstream of ATG8 family lipidation (MAP1LC3B, GABARAP, GABARAPL1) and autophagosome maturation. It forms functional complexes with ATG9, VMP1, and TMEM41B, and interacts directly with WIPI4, ATG12-ATG5-ATG16L1 conjugation machinery, thereby integrating core autophagy networks with lipid metabolism, endocytosis, and lysosomal degradation pathways.
In the colorectal cancer context, ATG2B knockout in HT29 cells provides a powerful platform to dissect autophagy’s dual roles in tumor suppression and promotion. Given the HT29 line’s mutant p53 status, this model is particularly suited to explore how loss of autophagy components influences cancer cell survival, metabolic adaptation, and therapeutic resistance. Disruption of ATG2B-mediated lipid transfer may impair autophagosome expansion, offering insights into tumor-specific vulnerabilities and the potential for targeting autophagy in p53-mutant cancers. Additionally, this model supports studies on lipid droplet dynamics and membrane trafficking in cancer, bridging fundamental cell biology with translational oncology.
Typical research applications include quantitative autophagy flux assays using bafilomycin A1 and LC3-II turnover measurement, immunofluorescence-based LC3 puncta quantification, and co-immunoprecipitation of ATG2B interactors. The polyclonal knockout population is also amenable to lipid transfer assays and functional readouts such as cell migration, invasion, and viability under nutrient stress or chemotherapy. Researchers can employ this model for drug resistance profiling, high-content screening of autophagy modulators, and mechanistic dissection of ATG2B-dependent signaling in colorectal cancer and beyond. For further technical details and ordering information, please contact Ascent Research.
