The BICD1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma line, engineered to disrupt BICD1 via CRISPR/Cas9-mediated gene disruption. This heterogeneous pool provides a loss-of-function model for studying BICD1-dependent intracellular processes without single-cell cloning.
The HT29 host cell line, isolated from a 44-year-old female colorectal adenocarcinoma patient, exhibits adherent epithelial morphology, mucin production, and a TP53 mutation (R273H). Widely utilized in colorectal cancer research, HT29 cells offer a well-characterized platform for investigating tumor biology and drug responses.
BICD1 encodes a dynein cargo adaptor that links Rab6 GTPase-positive Golgi-derived vesicles, endosomal compartments, and mRNA ribonucleoproteins to the dynein?Cdynactin motor complex, thereby enabling minus-end-directed transport along microtubules. This retrograde trafficking is critical for maintaining the integrity of the Golgi ribbon, proper endosomal positioning, and polarized secretion in epithelial cells. At the molecular level, BICD1 directly binds the dynein intermediate chain (DYNC1I1) and the dynactin subunit p50 (DCTN2), while its activity is modulated by upstream regulators including Rab6, RanBP2, and phosphorylation events. BICD1 also cooperates with the related adaptor BICDR-1 and functions within a larger pathway involving LIS1, NDE1, and NDEL1, which together coordinate cargo recognition and motor activation.
In the HT29 colorectal adenocarcinoma model, loss of BICD1 disrupts these critical transport processes, leading to potential Golgi fragmentation, endosomal mislocalization, and impaired exocytic and endocytic trafficking. Because HT29 cells are TP53 mutant and mucin-producing, this background allows investigation of how dynein-mediated transport intersects with tumor suppressor loss and secretory function. Defects in polarized intracellular transport are likely to compromise cell migration and invasion, making this knockout pool a valuable tool for studying mechanisms of colorectal cancer metastasis and resistance to therapies that target microtubule dynamics.
Researchers can leverage this polyclonal knockout population in diverse assays to probe BICD1 function. Immunofluorescence microscopy can visualize Golgi morphology (e.g., GM130, GRASP65) and endosome distribution (e.g., EEA1, Rab7); live-cell imaging using fluorescent organelle markers can track real-time trafficking; wound healing and transwell invasion assays can quantify migratory and invasive capacity; and biochemical approaches such as western blotting and co-immunoprecipitation can characterize protein expression and interactions. These experiments enable dissection of BICD1??s roles in dynein-mediated transport, colorectal cancer cell motility, and sensitivity to microtubule-targeting agents. For further information, please contact Ascent Research.
