The KATNAL1 Knockout HAP1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal cell population designed to disrupt the KATNAL1 gene in the HAP1 human cell line. This knockout model provides a loss-of-function system for investigating the cellular roles of KATNAL1, a microtubule-severing AAA ATPase critical for mitotic progression. The polyclonal format captures a range of genetic variants generated by CRISPR/Cas9-mediated disruption, offering a robust tool for functional studies without single-cell cloning.
The HAP1 cell line is a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid karyotype facilitates effective gene disruption, making it a preferred host for knockout screens and functional genomics. HAP1 cells retain key signaling pathways and cellular processes, including intact mitotic machinery, enabling the study of genes involved in fundamental biological processes such as cell division and cytoskeleton dynamics.
KATNAL1 encodes a microtubule-severing enzyme belonging to the AAA ATPase family, which is essential for spindle assembly and cytokinesis. The protein localizes to centrosomes and spindles, where it forms complexes with NDEL1 and LIS1, linking it to the dynein motor complex. Its activity is regulated by mitotic kinases: CDK1 phosphorylates KATNAL1 to modulate severing activity, while AURKA and PLK1 control its association with centrosomes. Downstream, KATNAL1-mediated severing generates microtubule fragments that influence NDEL1/LIS1/dynein-dependent transport and spindle organization. Disruption of KATNAL1 therefore impairs microtubule dynamics at centrosomes, leading to defective chromosome segregation.
In the HAP1 cellular context, loss of KATNAL1 disrupts the precise regulation of microtubule severing required for bipolar spindle formation and abscission. This results in mitotic delays and increased chromosome missegregation, phenotypes that can be monitored via time-lapse imaging or immunofluorescence detection of tubulin and mitotic markers. The model therefore provides a physiologically relevant system to examine how centrosomal microtubule defects contribute to genomic instability??a hallmark of cancer??as well as to explore mechanisms underlying KATNAL1-linked infertility and neurological disorders, where aberrant microtubule dynamics are implicated.
This polyclonal knockout pool is well-suited for a broad range of investigative applications, including cell cycle profiling by flow cytometry, analysis of mitotic structures via tubulin immunofluorescence, and biochemical characterization of KATNAL1-interacting complexes through co-immunoprecipitation with NDEL1 or LIS1. Functional assays such as time-lapse imaging of mitosis and proliferation measurements enable detailed dissection of spindle dynamics and division kinetics. The model is ideal for cancer research, drug target validation, genetic interaction screens, and mechanistic studies of microtubule regulation. For additional information or custom inquiries, please contact Ascent Research.
