Protein-protein interactions are key to inflammatory signaling and a multitude of proteins are pivotal to successful signal transduction in response to infection and inflammation. Some of these proteins demonstrate multifaceted mechanisms of control across multiple signaling pathways, which are often poorly understood. One such protein is macrophage migration inhibitory factor (MIF), a pleiotropic immunomodulatory protein that has been implicated in numerous inflammatory signaling pathways, including NF-kB and NLRP3 inflammasome activation. MIF is constitutively present in the cytosol and extracellular space, yet its mode of secretion is unclear. MIF is protective and pro-inflammatory in response to a range of bacterial pathogens, but potentially permissive to some viruses. Moreover, deletion or inhibition of MIF has been shown to decrease severity and/or susceptibility to a number of inflammatory and autoimmune diseases, including rheumatoid arthritis, lupus and inflammatory bowel disease. Here, we show that MIF inhibitors have varied effects on cytokine/chemokine release by human macrophage. While monoclonal antibodies and small molecule inhibitors of MIF have helped to decipher its role in mediating cytokine production and signal transduction, their specificity is questionable and there remains no consensus on the primary role of MIF. Thus, we are now utilising CRISPR-cas9 to tag endogenous MIF, either with a fluorescent protein or biotinylation enzyme, providing a pathway to investigate both transient and stable protein-protein interactions and cellular localisation. Covalently tagged MIF holds potential to decrease the number of non-MIF-specific hits currently seen with immune-precipitation and mass spectrometry. This approach will not only help us to uncover the mysteries of MIF but will provide a pipeline for immune protein interactome studies. Additionally, this project leverages CSIROs post-doc program and the Immune Resilience FSP to bridge the gap between traditional academic post-docs and industry projects while providing a personalised career development program.