The BOD1 Knockout Jurkat Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout population with targeted disruption of the BOD1 gene in the Jurkat T lymphoblast cell line. This loss-of-function model enables investigation of BOD1??s role in kinetochore function and chromosome segregation. The polyclonal format minimizes clonal bias and offers a heterogeneous cell pool suitable for functional studies.
Jurkat cells (clone E6-1) are a well-established human T-cell leukemia line commonly used to explore T-cell receptor signaling, apoptosis, and cell cycle regulation. Their suspension growth and robust proliferative capacity make them an ideal host for CRISPR-based gene disruption, particularly for studying mitosis in the context of hematologic malignancies.
BOD1 is a kinetochore-associated scaffold that recruits the PP2A-B56 phosphatase complex to kinetochores. There, PP2A-B56 dephosphorylates key substrates including the NDC80 complex and Ska complex, counteracting Aurora B kinase-mediated phosphorylation to stabilize kinetochore-microtubule attachments. BOD1 function is regulated by upstream kinases CDK1, PLK1, and Aurora B, which phosphorylate BOD1 to control its localization and activity. This regulatory circuit is critical for error correction and timely satisfaction of the spindle assembly checkpoint. Downstream, BOD1-dependent dephosphorylation events influence the NDC80 complex, Ska1, CENP-E, and other mitotic checkpoint proteins to ensure accurate chromosome segregation. Consequently, loss of BOD1 disrupts the kinetochore phospho-balance, causing chromosome misalignment, prolonged mitosis, and chromosomal instability.
In Jurkat T-cell leukemia cells, BOD1 knockout exacerbates mitotic errors and chromosomal instability, recapitulating a common feature of cancer. The polyclonal knockout pool is well-suited for dissecting how defective kinetochore regulation impacts cell cycle progression, aneuploidy, and leukemic cell proliferation. This model also facilitates screening of compounds that target mitotic regulators.
Researchers can apply these cells in live-cell imaging of mitotic dynamics, flow cytometry for cell cycle analysis, and immunofluorescence to visualize kinetochore-microtubule attachments. Western blotting enables detection of phosphorylated NDC80 and other mitotic phospho-targets, while functional assays including Annexin V apoptosis and CellTiter-Glo proliferation assays provide quantitative measures of viability and growth defects. The polyclonal population also facilitates screening for mitotic checkpoint defects using chemical inhibitors of Aurora B or PLK1, with readouts by high-content imaging. RT-qPCR can profile mitotic gene expression changes. For additional details, contact Ascent Research.
