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

Natacha Ipseiz, Magdalena A. Czubala, Valentina M.T. Bart, Luke Davies Orcid Logo, Robert H. Jenkins, Paul Brennan, Philip R. Taylor

Molecular Therapy - Methods & Clinical Development, Volume: 16, Pages: 21 - 31

Swansea University Author: Luke Davies Orcid Logo

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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...

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Published in: Molecular Therapy - Methods & Clinical Development
ISSN: 2329-0501
Published: Elsevier BV 2020
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URI: https://cronfa.swan.ac.uk/Record/cronfa61697
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spelling 2022-11-07T14:17:05.8939355 v2 61697 2022-10-31 Effective In Vivo Gene Modification in Mouse Tissue-Resident Peritoneal Macrophages by Intraperitoneal Delivery of Lentiviral Vectors ff080296775381560053d5e3a6e81745 0000-0001-7767-4060 Luke Davies Luke Davies true false 2022-10-31 BMS 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. Journal Article Molecular Therapy - Methods &amp; Clinical Development 16 21 31 Elsevier BV 2329-0501 13 3 2020 2020-03-13 10.1016/j.omtm.2019.10.004 COLLEGE NANME Biomedical Sciences COLLEGE CODE BMS Swansea University 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) 2022-11-07T14:17:05.8939355 2022-10-31T12:37:57.0375818 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine Natacha Ipseiz 1 Magdalena A. Czubala 2 Valentina M.T. Bart 3 Luke Davies 0000-0001-7767-4060 4 Robert H. Jenkins 5 Paul Brennan 6 Philip R. Taylor 7 61697__25671__38fbd3e6ee0447c289198a0230b85ddb.pdf 61697.pdf 2022-11-07T14:15:39.1351658 Output 2256227 application/pdf Version of Record true Copyright: 2019 The Authors.This is an open access article under the CC BY license true eng http://creativecommons.org/licenses/by/4.0/
title Effective In Vivo Gene Modification in Mouse Tissue-Resident Peritoneal Macrophages by Intraperitoneal Delivery of Lentiviral Vectors
spellingShingle Effective In Vivo Gene Modification in Mouse Tissue-Resident Peritoneal Macrophages by Intraperitoneal Delivery of Lentiviral Vectors
Luke Davies
title_short Effective In Vivo Gene Modification in Mouse Tissue-Resident Peritoneal Macrophages by Intraperitoneal Delivery of Lentiviral Vectors
title_full Effective In Vivo Gene Modification in Mouse Tissue-Resident Peritoneal Macrophages by Intraperitoneal Delivery of Lentiviral Vectors
title_fullStr Effective In Vivo Gene Modification in Mouse Tissue-Resident Peritoneal Macrophages by Intraperitoneal Delivery of Lentiviral Vectors
title_full_unstemmed Effective In Vivo Gene Modification in Mouse Tissue-Resident Peritoneal Macrophages by Intraperitoneal Delivery of Lentiviral Vectors
title_sort Effective In Vivo Gene Modification in Mouse Tissue-Resident Peritoneal Macrophages by Intraperitoneal Delivery of Lentiviral Vectors
author_id_str_mv ff080296775381560053d5e3a6e81745
author_id_fullname_str_mv ff080296775381560053d5e3a6e81745_***_Luke Davies
author Luke Davies
author2 Natacha Ipseiz
Magdalena A. Czubala
Valentina M.T. Bart
Luke Davies
Robert H. Jenkins
Paul Brennan
Philip R. Taylor
format Journal article
container_title Molecular Therapy - Methods &amp; Clinical Development
container_volume 16
container_start_page 21
publishDate 2020
institution Swansea University
issn 2329-0501
doi_str_mv 10.1016/j.omtm.2019.10.004
publisher Elsevier BV
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
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description 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.
published_date 2020-03-13T04:20:42Z
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