Journal article 626 views 81 downloads
Effective In Vivo Gene Modification in Mouse Tissue-Resident Peritoneal Macrophages by Intraperitoneal Delivery of Lentiviral Vectors
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Robert H. Jenkins,
Paul Brennan,
Philip R. Taylor
Molecular Therapy - Methods & Clinical Development, Volume: 16, Pages: 21 - 31
Swansea University Author:
Luke Davies
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Copyright: 2019 The Authors.This is an open access article under the CC BY license
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DOI (Published version): 10.1016/j.omtm.2019.10.004
Abstract
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 tar...
| Published in: | Molecular Therapy - Methods & Clinical Development |
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| ISSN: | 2329-0501 |
| Published: |
Elsevier BV
2020
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| Online Access: |
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| URI: | https://cronfa.swan.ac.uk/Record/cronfa61697 |
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| Abstract: |
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. |
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| College: |
Faculty of Medicine, Health and Life Sciences |
| Funders: |
P.R.T. is funded by a Wellcome Trust Investigator Award (107964/Z/15/Z), and this work was supported in part by the UK Dementia Research Institute. L.C.D. is funded by the Wellcome Trust (103973/Z/14/Z) |
| Start Page: |
21 |
| End Page: |
31 |