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Effective in vivo gene modification in mouse tissue-resident peritoneal macrophages by intraperitoneal delivery of lentiviral vectors

Ipseiz, Natacha ORCID:, Czubala, Magdalena A. ORCID:, Bart, Valentina M.T., Davies, Luke C., Jenkins, Robert H. ORCID:, Brennan, Paul ORCID: and Taylor, Philip R. ORCID: 2020. Effective in vivo gene modification in mouse tissue-resident peritoneal macrophages by intraperitoneal delivery of lentiviral vectors. Molecular Therapy - Methods and Clinical Development 16 , pp. 21-31. 10.1016/j.omtm.2019.10.004

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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 no options for tissue-specific targeting of macrophages. We have developed full protocols for in vivo productive infection (defined by stable transgene expression) of tissue-resident macrophages with lentiviral vectors, enabling RNA and protein overexpression, including expression of small RNA species such as shRNA, to knock down and modulate gene expression. These approaches allow robust infection of peritoneal tissue-resident macrophages without significant infection of other cell populations. They permit rapid functional study of macrophages in homeostatic and inflammatory settings, such as thioglycolate-induced peritonitis, while maintaining the cells in their physiological 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 background levels of inflammation induced by the virus. In summary, we present a quick and accessible protocol for the rapid assessment of gene function in peritoneal tissue-resident macrophages in vivo.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Publisher: Elsevier
ISSN: 2329-0501
Funders: Wellcome Trust
Date of First Compliant Deposit: 11 November 2019
Date of Acceptance: 14 October 2019
Last Modified: 25 Oct 2023 06:18

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