The GOLPH3 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting the GOLPH3 gene in Jurkat cells, providing a heterogeneous pool for robust investigation of GOLPH3-dependent processes. This loss-of-function model eliminates GOLPH3 protein expression, abolishing its roles in intracellular trafficking and signaling, and serves as a tool for dissecting GOLPH3 contributions in human T lymphoblast biology.
The host Jurkat cell line is an immortalized human T lymphoblastoid line from a 14-year-old with acute T cell leukemia, widely used for T cell signaling, apoptosis, and HIV studies. Its well-characterized signaling and rapid proliferation also render it a key model for T cell leukemia, enabling investigation of lymphomagenesis and drug resistance, making it an ideal background for studying GOLPH3’s oncogenic roles.
GOLPH3 encodes a Golgi peripheral membrane protein that binds phosphatidylinositol-4-phosphate (PtdIns(4)P) and the motor protein MYO18A to facilitate Golgi-to-plasma membrane trafficking and RhoA-dependent Golgi dispersal. This activity enhances secretion and glycosylation of growth factor receptors. GOLPH3 activates mTORC1 by interacting with VPS35, a component of the retromer complex, leading to phosphorylation of S6K1 and 4EBP1. It is transcriptionally induced by MYC and stimulated by PI3K/AKT signaling, and it interacts with DNA-PK to inhibit DNA damage-induced apoptosis, thereby promoting oncogenesis in multiple malignancies.
In Jurkat T lymphoblasts, GOLPH3 knockout cells enable dissection of mTORC1-driven growth and Golgi-mediated secretion, particularly glycosylation of T cell receptor components and cytokine receptors. This allows the study of how GOLPH3 loss impacts signal transduction, apoptotic thresholds, and immune synapse formation, offering a model for hematologic malignancies where Golgi dysregulation contributes to transformation.
Research applications include Western blotting for mTORC1 targets (phospho-S6K1, phospho-4EBP1), flow cytometry for surface glycosylation and Annexin V apoptosis assays, co-immunoprecipitation to assess GOLPH3-VPS35/MYO18A interactions, and cell migration and chemosensitivity assays. The cells are also suitable for siRNA rescue experiments and high-content screens against the Golgi-mTOR axis. For additional technical support, please contact Ascent Research.
