The ANAPC7 Knockout Jurkat Polyclonal Cells are a human T-lymphoblast suspension model with CRISPR/Cas9-mediated disruption of the ANAPC7 gene. This polyclonal knockout cell population targets the anaphase-promoting complex/cyclosome (APC/C) core subunit ANAPC7 via genome editing, avoiding clonal selection biases. The loss-of-function model retains Jurkat growth properties and enables robust APC/C pathway studies.
Derived from a 14-year-old male with acute T-cell leukemia, the Jurkat host cell line is widely used for T-cell signaling, apoptosis, and leukemia research. These T-lymphoblasts proliferate rapidly in suspension, making them ideal for cell cycle and cancer biology investigations. Combined with ANAPC7 polyclonal knockout, this system provides a physiologically relevant platform to study mitotic regulation in a T-cell malignancy context.
ANAPC7 encodes a core APC/C subunit, an E3 ubiquitin ligase that targets mitotic regulators for proteasomal degradation. With coactivators CDC20 and CDH1, APC/C ubiquitinates Cyclin B1 (CCNB1), Cyclin A2 (CCNA2), securin (PTTG1), and NEK2A, enabling chromosome segregation and mitotic exit. Upstream kinases CDK1 and PLK1 activate APC/C, while spindle assembly checkpoint proteins MAD2L1 and BUB1B inhibit CDC20. E2 enzymes UBE2C and UBE2S cooperate with APC/C for ubiquitin chain formation. ANAPC7 disruption impairs complex assembly, causing cyclin and securin accumulation, mitotic arrest, and chromosomal instability.
In Jurkat T-cell leukemia, ANAPC7 knockout perturbs cell cycle progression, reflecting aneuploidy-associated mitotic defects. The polyclonal population preserves editing heterogeneity, reducing clonal adaptation artifacts and revealing varied loss-of-function phenotypes. This model is valuable for studying APC/C dysfunction in leukemogenesis, given T-lymphoblasts’ high mitotic rate and reliance on proteasomal degradation. It enables assessment of proliferation, chromosomal fidelity, and sensitivity to mitotic checkpoint inhibitors in a leukemic background.
Applications include flow cytometric cell cycle profiling, Western blotting for cyclins and securin, and immunofluorescence of mitotic spindles. Co-immunoprecipitation detects APC/C complex interactions, while ubiquitination assays evaluate E3 ligase activity. Mitotic synchronization and release protocols permit tracking of mitotic progression. The model supports target validation, drug screening for aneuploidy-driven cancers, and USP pathway studies in T-cell leukemia. For further details or ordering, contact Ascent Research.
