The BOD1 Knockout HT29 Polyclonal Cells product provides a CRISPR/Cas9?mediated loss?of?function model for the kinetochore protein BOD1 in a colorectal adenocarcinoma background. This polyclonal knockout cell population was generated by introducing targeted gene disruptions in the BOD1 locus of HT29 cells, resulting in a heterogeneous pool of knockout alleles. The product is intended for use in mitotic studies, chromosome instability research, and anticancer drug testing.
The host HT29 cell line is a well?characterized human epithelial colorectal adenocarcinoma model originally isolated from a primary tumor of a 44?year?old female patient. HT29 cells are extensively employed in intestinal epithelial biology, drug permeability and absorption studies, and cancer research owing to their ability to differentiate and produce mucins under appropriate culture conditions. The parental line retains key features of colorectal tumorigenesis, making it a relevant background for investigating genes involved in chromosomal instability and mitotic dysfunction.
BOD1 (Biorientation of chromosomes in cell division 1) is a kinetochore?associated protein that ensures faithful chromosome segregation during mitosis. It is recruited to kinetochores where it serves as a platform for the kinesin?7 motor CENP?E by interacting with the NDC80 complex and the KMN network (KNL1/Mis12/Ndc80). This recruitment stabilizes kinetochore?microtubule attachments, promoting chromosome congression to the metaphase plate. BOD1 function is tightly controlled by phosphorylation from CDK1, Aurora A, Aurora B, and PLK1. Knockout of BOD1 prevents CENP?E loading, resulting in defective microtubule capture, chromosome misalignment, and sustained activation of the spindle assembly checkpoint through downstream effectors BUB1, BUBR1, and MAD2.
In the HT29 colorectal adenocarcinoma background, BOD1 knockout provides a physiologically relevant system to study the interplay between mitotic errors and colorectal cancer biology. HT29 cells exhibit moderate chromosomal instability and are responsive to alterations in kinetochore function, making them suitable for examining how BOD1 loss exacerbates aneuploidy, affects cell cycle progression, and influences sensitivity to anti?mitotic agents. This polyclonal population mirrors the heterogeneity present in tumors, offering a robust tool for investigating the consequences of BOD1 deficiency on chromosome segregation fidelity, ploidy changes, and tumor cell proliferation.
Researchers can use these knockout cells in immunofluorescence microscopy to assess kinetochore alignment and spindle morphology, live?cell imaging for chromosome dynamics, and Western blotting for mitotic markers such as phospho?histone H3 and cyclin B1. Flow cytometry enables cell cycle and ploidy analysis, while proliferation assays (MTT, BrdU) and drug sensitivity screens with taxanes or Aurora kinase inhibitors provide functional insights. This model is a valuable tool for studying chromosome instability and testing anti?mitotic therapies in colorectal cancer. For technical support, please contact Ascent Research.
