The BRSK1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HAP1 cell line, offering a loss-of-function model for the brain-specific serine/threonine kinase BRSK1. This product comprises a heterogeneous pool of cells with targeted gene disruptions, enabling pooled functional studies without clonal selection artifacts. The polyclonal format is particularly suited for genetic screens and assays where population-level phenotypes are informative, and it avoids the limitations inherent to single-cell-derived clones. It provides a robust platform for investigating neuronal polarization, microtubule dynamics, and AMPK-related signaling in a cancer-relevant, suspension-adapted host.
The HAP1 host line is a suspension-adapted near-haploid human leukemic cell line derived from KBM-7, originally from a chronic myeloid leukemia patient. Its near-haploid karyotype??most chromosomes single copy??enables recessive phenotypes with single-allele disruption, ideal for haploid genetic screens. The leukemic origin provides context for cancer proliferation studies, including leukemia-specific signaling vulnerabilities, and suspension growth facilitates high-throughput automated screening workflows.
BRSK1 belongs to the AMPK-related kinase family and is activated by LKB1 (STK11) in complex with STRADA and CAB39. It directly phosphorylates Tau and MAP2 to regulate microtubule stability, and phosphorylates ??-tubulin at centrosomes for centrosome duplication control. Interactions with the related kinase BRSK2 and scaffold 14-3-3 (YWHAE) integrate BRSK1 into AMPK and mTOR signaling networks, influencing neuronal polarization, cell cycle progression, and synaptic function via downstream target SYNGAP1.
In HAP1 cells, BRSK1 knockout disrupts LKB1-mediated signaling, enabling detailed study of microtubule regulation and cell division fidelity. The haploid background enhances phenotypic penetrance and simplifies genetic modifier screens. BRSK1??s role in centrosome biology and mitotic progression makes this model particularly relevant for investigating chromosome instability in leukemia. The polyclonal population mimics tumor heterogeneity, enhancing translational cancer research.
Representative applications include western blotting for phospho-Tau and phospho-??-tubulin, immunofluorescence for microtubule organization, flow cytometric cell cycle profiling, and migration/invasion assays. The haploid background supports genome-wide CRISPR screens to identify genetic interactions such as synthetic lethal partners or resistance modifiers. Phospho-signaling arrays and co-immunoprecipitation experiments can further map altered signaling networks. For comprehensive technical data, please contact Ascent Research.
