The GOLPH3 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the GOLPH3 gene in a near-haploid human cell background. This product provides a heterogeneous pool of edited cells, enabling robust assessment of gene function without clonal selection artifacts. The polyclonal format is particularly suited for pooled phenotypic screens and pathway dissection experiments that require a population-level representation of gene disruption effects. By employing CRISPR/Cas9-mediated gene disruption, the GOLPH3 locus is targeted to abrogate protein expression, creating a versatile knockout model for cancer and cell biology research.
The host cell line, HAP1, is a near-haploid myeloid leukemia cell line derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid karyotype simplifies genetic analysis by reducing gene copy number to a single allele for most chromosomes, making it an ideal platform for functional genomics and knockout-based investigations. HAP1 cells retain key characteristics of leukemic cells and exhibit robust growth in culture, facilitating high-throughput genetic screens and biochemical assays. This genetic simplicity allows for unambiguous interpretation of gene knockout phenotypes, particularly in signaling and trafficking pathways.
GOLPH3 (Golgi phosphoprotein 3) is an oncogenic protein localized to the trans-Golgi network, where it regulates Golgi-to-plasma membrane trafficking and glycosylation. Mechanistically, GOLPH3 interacts with VPS35 to enhance mTORC1 activation, leading to increased phosphorylation of downstream effectors including AKT, S6K1, and 4E-BP1. This signaling is further modulated by upstream regulators such as MYC and PI3K/AKT, and GOLPH3 also participates in DNA damage-induced Golgi dispersal via interactions with DNA-PKcs and MYO18A. Through these interactions, GOLPH3 promotes cell proliferation, migration, and survival, establishing its central role in oncogenic signaling networks.
In the near-haploid HAP1 background, knockout of GOLPH3 provides a powerful model to dissect its contributions to mTOR signaling and Golgi-dependent trafficking without compensatory effects from a second allele. The leukemic origin of HAP1 cells offers a relevant context for studying GOLPH3??s role in cancer, particularly in hematological malignancies and solid tumors where GOLPH3 is frequently overexpressed. This model enables precise interrogation of GOLPH3-dependent phenotypes, including altered glycosylation patterns and deregulated mTORC1 activity, which are critical for understanding tumorigenesis and identifying therapeutic vulnerabilities.
Researchers can utilize these polyclonal knockout cells in a variety of experimental applications, including Western blotting to confirm loss of GOLPH3 and downstream phospho-AKT levels, proliferation and migration assays to assess functional consequences, and immunofluorescence staining of Golgi markers to examine trafficking defects. Co-immunoprecipitation studies can be performed to validate interactions with VPS35 or DNA-PKcs, while cell cycle analysis and glycosylation profiling provide deeper insight into GOLPH3-mediated cellular processes. This product is an invaluable tool for cancer research, signal transduction studies, and drug target identification. For further information, please contact Ascent Research.
