Effective In?Vivo Gene Modification in Mouse Tissue-Resident Peritoneal Macrophages by Intraperitoneal Delivery of Lentiviral Vectors

摘要

Tissue-resident macrophages exhibit specialized phenotypes dependent on their in vivo physiological niche. Investigation of their function often relies upon complex whole mouse transgenic studies. While some appropriate lineage-associated promoters exist, there are nooptions for tissue-specific targeting of macrophages. We have developed full protocols for in vivo productive infection (defined by stabletransgene expression) of tissue-resident macrophages with lentiviral vectors, enabling RNA and protein overexpression, includingexpression of small RNA species such as shRNA, to knock down and modulate gene expression. These approaches allow robust infectionof peritoneal tissue-resident macrophages without significant infection of other cell populations. They permit rapid functional study ofmacrophages in homeostatic and inflammatory settings, such as thioglycolate-induced peritonitis, while maintaining the cells in theirphysiological context. Here we provide detailed protocols for the whole workflow: viral production, purification, and quality control;safety considerations for administration of the virus to mice; and assessment of in vivo transduction efficiency and the low backgroundlevels of inflammation induced by the virus. In summary, we present a quick and accessible protocol for the rapid assessment of genefunction in peritoneal tissue-resident macrophages in vivo.