The AGTPBP1 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal population with targeted disruption of AGTPBP1 in a human near-haploid background. This heterogeneous pool enables robust loss-of-function studies without clonal selection bias. AGTPBP1 encodes a deglutamylating carboxypeptidase critical for removing polyglutamate chains from tubulin, thereby modulating microtubule dynamics and protein function. The knockout model is optimally suited for high-throughput functional genomics, drug screening, and detailed mechanistic investigations of tubulin post-translational modifications.
HAP1 cells are a chronic myeloid leukemia (CML)-derived near-haploid cell line originating from the KBM-7 parental line. Their haploid karyotype simplifies CRISPR/Cas9-mediated gene disruption, as a single targeted allele suffices for complete knockout. Widely employed for genetic screens, target validation, and disease modeling, HAP1 cells retain CML-associated molecular features, making them particularly relevant for cancer cell biology studies and investigations of microtubule-targeting agents in a leukemia context.
AGTPBP1 functions as a cytosolic carboxypeptidase that selectively removes polyglutamate side chains from the C-terminal tails of tubulin, counterbalancing tubulin polyglutamylases. This deglutamylation event regulates microtubule stability and governs the binding of microtubule-associated proteins (MAPs) such as tau and MAP1A. In the knockout state, hyperglutamylation predominates, leading to disrupted microtubule network integrity, mitotic spindle defects, and impaired intracellular transport. Core pathway components include tubulin, polyglutamylase enzymes (e.g., TTLL family members), AGTPBP1, and downstream MAP effectors, with pathway activity modulated by cell cycle-dependent signals.
In HAP1 cells, AGTPBP1 knockout leads to pronounced tubulin hyperglutamylation, providing a clean genetic system to dissect microtubule-related phenotypes. The haploid state eliminates potential compensatory expression from a second allele, ensuring a complete loss of deglutamylase function. This model is sensitive for detecting alterations in microtubule stability, cell cycle progression, and apoptotic responses. It is particularly valuable for modeling neurodegenerative conditions such as cerebellar atrophy linked to AGTPBP1 mutations and for investigating how tubulin post-translational modifications influence cancer cell sensitivity to microtubule-targeting chemotherapeutics.
These polyclonal knockout cells are compatible with western blotting for tubulin glutamylation, immunofluorescence microscopy, microtubule stability assays, RT-qPCR for knockout confirmation, flow cytometric cell cycle analysis, and drug sensitivity profiling with microtubule-directed agents. The polyclonal format captures multiple independent editing events, enhancing statistical rigor. For further technical information, please contact Ascent Research.
