The DTX3L Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population specifically engineered for loss-of-function analysis of the DTX3L gene in the human HT29 colorectal adenocarcinoma cell line. This polyclonal format provides a heterogeneous pool of cells with disrupted DTX3L, avoiding clonal selection artifacts and enabling robust loss-of-function studies.
HT29 cells are an adherent, epithelial cell line derived from human colorectal adenocarcinoma. They serve as a widely used model for studying colorectal cancer biology, intestinal epithelial function, and oncogenic signaling pathways. Their relevance to colorectal tumorigenesis makes them suitable for investigating genes involved in proliferation, differentiation, and immune interaction.
DTX3L functions as an E3 ubiquitin ligase that ubiquitinates Notch receptors, targeting them for proteasomal degradation and attenuating Notch signaling. It also partners with PARP9 to enhance interferon signaling by activating IRF3 and IRF7 phosphorylation and promoting NF-??B p65 activity. Upstream, interferons (IFN-??, IFN-??) induce DTX3L via JAK1/TYK2 kinases and STAT1/STAT2 transcription factors. Notch ligands DLL1, DLL4, JAG1, JAG2 trigger receptor processing, and DTX3L ubiquitinates the resulting Notch intracellular domain (NICD). Downstream effects include modulation of ISGs such as IRF1 and MX1, and Notch targets like HES1.
In colorectal cancer, aberrant Notch and interferon pathways contribute to tumor progression and immune evasion. DTX3L’s dual role as a Notch repressor and interferon amplifier positions it as a key regulator of these pathways. Knocking out DTX3L in HT29 cells enables dissection of tumor-intrinsic immune signaling and Notch-dependent differentiation, providing insights into colorectal cancer mechanisms and potential therapeutic targets.
Applications include western blotting for NICD, RT-qPCR of ISGs (IRF1, MX1), immunofluorescence for NF-??B nuclear translocation, flow cytometry for surface Notch1, and co-immunoprecipitation of DTX3L-PARP9. Functional assays such as apoptosis, migration/invasion, and colony formation can assess cellular phenotypes. Transcriptome analysis by RNA-seq upon interferon stimulation reveals global expression changes. These polyclonal knockout cells are a versatile and highly applicable tool for colorectal cancer and innate immunity research. For further information, please contact Ascent Research.
