Generation of Stable Cell Lines Expressing Golgi Reassembly Stacking Proteins (GRASPs) by Viral Transduction

Stable cell linesStable cell lines that express a gene of specific interest provide an advantage over transient gene expression by reducing variations in transfectionTransfection efficiency between experiments, sustaining expression for long-term studies, and controlling expression levels in particular if a clonal population is selected. Transient transfectionTransfection requires introduction of an exogenous gene into host cells via typically harsh chemicals or conditions that permeabilize the cell membraneMembranes, which does not normally integrate into the target cell genome. Here, we describe the methodMethods of using retroviral transductionTransductions to stably express Golgi proteinsGolgi proteins fused to a promiscuous biotin ligase (TurboIDTurboID) in HeLa cells, thus creating cell lines that can be leveraged in studies of the proximome/interactome. We also demonstrate a similar protocol for stable expression of a Golgi proteinGolgi proteins fused to a fluorescent tag via lentiviral transductionTransductions. These methodsMethods can be further adapted to establish other cell lines with different sub-cellular markersMarkers or fusion tags. Viral transductionTransductions is a convenient methodMethods to create stable cell linesStable cell lines in cell-based studies.