The KATNA1 Knockout HT29 Polyclonal Cells product provides a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population derived from the HT29 human colorectal adenocarcinoma line, targeting the KATNA1 locus. This polyclonal knockout model is generated without single-cell cloning, preserving a heterogeneous pool of edited alleles that collectively ablate KATNA1 function. The product is designed for loss-of-function studies investigating microtubule-severing ATPase activity in an intestinal epithelial context, enabling robust interrogation of mitotic spindle regulation and cytoskeletal dynamics without clonal selection bias.
HT29 cells are an adherent epithelial line isolated from a human colorectal adenocarcinoma, widely employed as a model for colorectal cancer biology. These cells retain characteristic epithelial morphology and harbor mutations in key oncogenic and tumor-suppressor pathways, including APC and TP53, making them a relevant system for studying transformation-associated processes. Their adherent growth and well-differentiated features facilitate high-resolution imaging of subcellular structures, such as the mitotic spindle and microtubule networks, and they support quantification of proliferation, migration, and invasion in 2D and 3D culture formats.
KATNA1 encodes the p60 catalytic subunit of the heterodimeric microtubule-severing AAA ATPase katanin. Katanin activity is regulated by phosphorylation through CDK1 kinase and Aurora A kinase, especially during mitotic entry, and is essential for severing microtubules into shorter fragments. KATNA1 forms a functional complex with the regulatory p80 subunit KATNB1, and this complex directly interacts with tubulin heterodimers and microtubule-associated proteins on spindle microtubules. Active katanin promotes microtubule depolymerization and contributes to mitotic spindle turnover, impacting spindle assembly, chromosome segregation, and ciliogenesis. Disruption of KATNA1 therefore compromises the controlled severing required for dynamic microtubule remodeling.
In HT29 colorectal cancer cells, loss of KATNA1-mediated microtubule severing impairs mitotic spindle organization and dynamics, leading to defects in chromosome alignment and segregation, prolonged mitosis, and reduced proliferative capacity. Additionally, microtubule network alterations may hinder directed cell migration and invasion, processes central to metastatic progression. This knockout model is therefore highly relevant for dissecting the contribution of microtubule severing to colorectal tumor cell behavior and for evaluating katanin as a therapeutic vulnerability in cancer.
Researchers can utilize this product for mechanistic studies using immunofluorescence microscopy to visualize spindle defects and acetylated tubulin stabilization, western blotting to confirm KATNA1 loss and downstream tubulin modification, flow cytometry for cell cycle arrest analysis, and colony formation or migration assays to assess tumorigenic phenotypes. These applications support investigation of microtubule-severing dynamics in mitosis, katanin-dependent regulation of intestinal epithelial division, and screening of anti-mitotic compounds targeting katanin function. For further details or to discuss custom applications, please contact Ascent Research.
