The GOLIM4 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting GOLIM4 in HAP1 cells. This mixed pool of edited cells, generated via CRISPR/Cas9-mediated gene disruption, provides a loss-of-function model for studying Golgi biology without clonal expansion. The polyclonal format captures diverse editing outcomes, enabling robust functional assays and pooled screens.
HAP1 is a near-haploid cell line derived from chronic myelogenous leukemia KBM-7 cells, maintaining haploidy for all chromosomes except chromosome 8. This genetic simplicity allows efficient homozygous knockout generation with a single guide RNA, making it ideal for genetic screens. The adherent HAP1 background retains cancer-relevant pathways, offering a relevant model for investigating Golgi function in a CML context.
GOLIM4 is a Golgi integral membrane protein essential for Golgi stack architecture and vesicular transport. It interacts with Golgi matrix proteins such as golgins, and operates within a network including GRASP55/GRASP65, COPI coatomer, and cis-Golgi matrix proteins. GOLIM4 disruption impairs Golgi integrity, leading to defective protein sorting, glycosylation, and secretion. While its upstream regulators and downstream effectors are unknown, GOLIM4??s role in glycoprotein processing and trafficking makes it a critical component of Golgi organization.
The HAP1 polyclonal knockout model is highly advantageous for Golgi research due to the host??s near-haploid genome, which simplifies genotype-phenotype correlations. The heterogeneous knockout pool facilitates investigation of phenotypic variability in protein trafficking and glycosylation in a cancer background, where Golgi dysfunction is implicated. This system supports combinatorial studies with other Golgi-related genes and synthetic lethality screens.
Applications include immunofluorescence for Golgi markers (GM130, Giantin), western blotting for glycoproteins (LAMP1, LAMP2), flow cytometry for surface receptors, and secretion assays (luciferase reporter). Proliferation assays can assess effects on cell growth. These cells serve cancer biology, functional genomics, and drug target validation. For further information, please contact Ascent Research.
