The GORASP1 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the human GORASP1 (Golgi reassembly stacking protein 1, also known as GRASP65) gene has been disrupted. This loss-of-function model is designed for researchers investigating the structural and regulatory roles of GORASP1 in the Golgi apparatus, providing a versatile tool for studying Golgi biology, secretion pathways, and related disease mechanisms.
The HAP1 cell line is a near-haploid human cell line derived from KBM-7 chronic myeloid leukemia cells, exhibiting fibroblast-like morphology and a near-haploid karyotype. Its genetic simplicity and stable growth characteristics make it an excellent host for knockout studies, enabling clear genotype?Cphenotype correlations and high-throughput genetic and pharmacological screens.
GORASP1 is a key Golgi stacking protein that forms a complex with GM130 (GOLGA2) to maintain cisternal stacking and ribbon integrity. Phosphorylation by CDK1/CCNB1, PLK1, or ERK1/2 (MAPK3/MAPK1) downstream of RAS?CRAF?CMEK disrupts this interaction, triggering Golgi unstacking and fragmentation. This process modulates secretion of cargo such as cytokines and integrins, and impacts integrin trafficking, cell adhesion, and migration. GORASP1 also interacts with GRASP55, p115 (USO1), Rab GTPases, golgins, and COPI coat components, placing it at the center of Golgi dynamics and transport. It also regulates autophagy and unconventional secretion pathways.
In the HAP1 near-haploid background, the single-allele disruption of GORASP1 eliminates genetic redundancy, providing a clean model to dissect kinase-substrate relationships (e.g., CDK1?CGORASP1, ERK?CGORASP1) involved in mitotic Golgi fragmentation and reassembly. Moreover, the HAP1 system allows direct biochemical analysis of GORASP1?CGM130 interactions and the effects of kinase inhibitors, enabling detailed mechanistic studies. This system is ideal for correlating biochemical changes with functional outcomes and for screening compounds that affect Golgi morphology.
Typical applications include immunofluorescence staining for Golgi markers (GM130, Giantin), electron microscopy for ultrastructure, secretion assays, wound healing and transwell migration assays, phospho-specific western blotting for GORASP1, and co-immunoprecipitation with GM130. Further, these cells can be utilized in drug discovery efforts targeting Golgi function or cancer cell migration. The polyclonal nature ensures a population-level representation of knockout effects while maintaining high reproducibility. For additional information or to explore customized applications, contact Ascent Research.
