The ANKRD50 Knockout HT29 Polyclonal Cells product provides a versatile CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line. This heterogeneous pool encompasses diverse gene-disruption alleles at the ANKRD50 locus, enabling loss-of-function studies immediately upon thawing without the need for clonal isolation or single-cell expansion. The cells are supplied as a frozen vial of early-passage stock, ensuring robust viability, stable growth, and reproducible experimental outcomes across multiple assays.
The HT29 host line originates from a primary colon adenocarcinoma resected from a 44-year-old female and has become a widely employed model for colorectal cancer research. HT29 cells carry defined mutations in the adenomatous polyposis coli (APC) and TP53 tumor suppressor genes, while retaining wild-type KRAS, which drives constitutive activation of the Wnt/??-catenin signaling pathway. These adherent epithelial cells differentiate into polarized monolayers with functional tight junctions, making them particularly suitable for investigating intestinal barrier integrity, drug absorption and transport, and tumor cell biology in a well-characterized genetic background.
ANKRD50 (Ankyrin Repeat Domain 50) is predicted to encode a scaffold protein featuring ankyrin repeat motifs that mediate protein-protein interactions, with localization studies suggesting centrosomal and ciliary enrichment. It is hypothesized to participate in centrosome duplication and ciliogenesis, potentially interacting with centrosomal components such as CEP152, CPAP, and STIL. ANKRD50 has been linked to the Wnt/??-catenin signaling axis through transcriptional effector TCF4 (TCF7L2), and its disruption may influence downstream targets involved in cell cycle progression and cilia assembly. Although its upstream regulators remain unknown, the protein??s domain architecture and interaction network implicate it in centrosome regulation and signal integration.
Disrupting ANKRD50 in the HT29 background creates a unique system to study the interplay between centrosome biology and colorectal cancer pathogenesis. Since HT29 cells exhibit constitutive Wnt pathway activation due to APC mutation, adding ANKRD50 knockout allows dissection of how centrosome-associated scaffolding influences Wnt-driven proliferation, migration, and colony-forming capacity. Moreover, given ANKRD50??s association with neurodevelopmental disorders such as intellectual disability and microcephaly, this model offers a cancer-relevant platform for exploring ciliopathy mechanisms. The knockout can reveal potential roles in maintaining genomic stability or modulating cilia-dependent signaling that impacts tumor cell behavior.
Researchers can employ this polyclonal knockout cell pool in a variety of experimental settings: western blotting and RT-qPCR confirm ANKRD50 depletion; immunofluorescence with anti-CEP152 or anti-CPAP antibodies visualizes centrosome morphology; cell proliferation, cell cycle flow cytometry, and colony formation assays measure growth phenotypes; migration assays assess metastatic traits; and serum-starvation-induced ciliogenesis assays test cilia formation. Indel detection by sequencing validates gene editing. These applications support functional characterization of ANKRD50, investigation of centrosome and cilia biology, and modeling of colorectal cancer pathogenesis. For further information, please contact Ascent Research.
