GOLGA4 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for investigating the role of GOLGA4 in Golgi trafficking and leukemic cell biology. This product contains a heterogeneous pool of HAP1 cells harboring gene disruption at the GOLGA4 locus, providing a loss-of-function model to dissect GOLGA4-dependent processes without the clonal artifacts associated with single-cell-derived lines. The polyclonal format enables robust and reproducible study of vesicle tethering and membrane trafficking mechanisms.
The HAP1 parental line is a near-haploid adherent cell line derived from the KBM-7 chronic myeloid leukemia (CML) patient isolate. HAP1 cells retain expression of the BCR-ABL fusion protein, a hallmark driver of CML pathogenesis, making them a pertinent hematopoietic progenitor model for studying Golgi biology in the context of oncogenic signaling. The near-haploid genome facilitates straightforward interpretation of gene-editing outcomes and permits efficient lentiviral transduction, while the adherent nature enables convenient imaging-based assays.
GOLGA4 encodes a golgin family coiled-coil protein that serves as a tethering factor at the trans-Golgi network (TGN). It is activated by the small GTPases ARL1 and RAB1, which recruit GOLGA4 to TGN membranes where it captures Golgi-derived vesicles targeted to the plasma membrane. GOLGA4 subsequently interacts with components of the COPI complex, the GRASP55 stacking protein, and TMF1 to organize cargo sorting and vesicle formation. It functions upstream of SNARE-mediated fusion events that deliver plasma membrane proteins and secreted factors. Deletion of GOLGA4 disrupts Golgi architecture, impairs retrograde and anterograde trafficking, and alters the surface expression of receptors and adhesion molecules.
In the HAP1 CML background, GOLGA4 loss-of-function provides a unique tool to examine how Golgi-dependent trafficking intersects with BCR-ABL-driven signaling. Aberrant Golgi organization is increasingly recognized in cancer cell migration, drug resistance, and secretory pathway dysregulation. GOLGA4 knockout in HAP1 cells may compromise the delivery of integrins, growth factor receptors, and secreted autocrine factors that support leukemic cell survival and motility, thus offering a model to study these processes. Additionally, the polyclonal population reduces the risk of off-target site homozygosity, enhancing the reliability of comparative phenotypic screens.
Researchers can employ this polyclonal knockout pool in a variety of assays, including immunofluorescence-based analysis of Golgi morphology, Western blotting to confirm loss of GOLGA4 protein, and RT-qPCR for mRNA quantification. Functional studies may utilize the VSV-G trafficking assay to measure secretory pathway kinetics, wound healing migration assays to assess cell motility, and flow cytometry to profile cell surface marker abundance. These cells are well-suited for drug sensitivity screens that probe the role of Golgi trafficking in chemoresistance and for co-culture experiments examining tumor?Cstromal interactions. For additional technical details and availability, please contact Ascent Research.
