This product is a CRISPR/Cas9-edited polyclonal knockout cell population targeting the KIF2A gene in the HT29 human colorectal adenocarcinoma cell line. The targeted disruption of the KIF2A locus generates a loss-of-function model within a heterogeneous cell pool, allowing researchers to investigate the functional consequences of KIF2A deficiency without clonal selection artifacts. The polyclonal format preserves genomic diversity while ensuring robust depletion of KIF2A protein expression, making it suitable for population-level functional assays in cancer cell biology and microtubule dynamics research.
HT29 cells are an adherent epithelial cell line originally isolated in 1964 from a primary colorectal adenocarcinoma of a 44-year-old female patient. They serve as a widely employed model for intestinal epithelial biology, colorectal cancer progression, and drug absorption studies due to their ability to form polarized monolayers and express relevant differentiation markers. The cells retain key oncogenic mutations, including APC, TP53, and KRAS, which contribute to their transformed phenotype, making them a representative system for studying molecular mechanisms in colorectal tumorigenesis.
KIF2A encodes a kinesin-13 family microtubule depolymerase that catalyzes ATP-dependent microtubule disassembly from both plus and minus ends, playing an essential role in mitotic spindle assembly, chromosome segregation, and axonal pruning. The enzymatic activity of KIF2A is tightly regulated by upstream kinases: Aurora A phosphorylates KIF2A to control spindle pole focusing, while CDK1/Cyclin B and PLK1 modulate its depolymerase function during mitosis. KIF2A directly interacts with microtubules and plus-end tracking proteins such as EB1, TIP150, and CLIP-170 to spatially regulate depolymerization. Loss of KIF2A disrupts a critical signaling axis involving Aurora A, PLK1, and the NDC80 complex, leading to destabilized kinetochore-microtubule attachments and prolonged mitosis.
In the context of HT29 colorectal cancer cells, disruption of KIF2A renders this model particularly informative for studying mitotic vulnerabilities and cytoskeletal regulation in transformed epithelial cells. KIF2A knockout is expected to impair microtubule dynamics, resulting in mitotic spindle abnormalities, chromosome misalignment, and altered cell cycle progression. These defects can influence tumor cell proliferation and migration, providing a relevant platform to dissect the contribution of microtubule-regulating enzymes to colorectal cancer aggressiveness and to validate KIF2A as a potential therapeutic target in malignancies characterized by chromosomal instability.
This polyclonal knockout cell population is suited for a broad range of experimental applications, including western blotting to confirm KIF2A depletion, immunofluorescence microscopy to visualize mitotic spindle morphology, and flow cytometry for cell cycle profiling. Functional assays such as colony formation, proliferation kinetics, and migration assays can be employed to assess the impact on tumor cell behavior, while microtubule regrowth assays enable direct measurement of depolymerase activity. Researchers may also combine this model with pharmacological inhibitors of Aurora A or PLK1 to explore synthetic lethality or drug resistance mechanisms. For additional product details or technical support, please contact Ascent Research.
