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Cas9(+) conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond
Macrophages play critical roles in immunity, development, tissue repair, and cancer, but studies of their function have been hampered by poorly-differentiated tumor cell lines and genetically-intractable primary cells. Here we report a facile system for genome editing in non-transformed macrophages...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6579556/ https://www.ncbi.nlm.nih.gov/pubmed/31204998 http://dx.doi.org/10.7554/eLife.45957 |
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author | Roberts, Allison W Popov, Lauren M Mitchell, Gabriel Ching, Krystal L Licht, Daniel J Golovkine, Guillaume Barton, Gregory M Cox, Jeffery S |
author_facet | Roberts, Allison W Popov, Lauren M Mitchell, Gabriel Ching, Krystal L Licht, Daniel J Golovkine, Guillaume Barton, Gregory M Cox, Jeffery S |
author_sort | Roberts, Allison W |
collection | PubMed |
description | Macrophages play critical roles in immunity, development, tissue repair, and cancer, but studies of their function have been hampered by poorly-differentiated tumor cell lines and genetically-intractable primary cells. Here we report a facile system for genome editing in non-transformed macrophages by differentiating ER-Hoxb8 myeloid progenitors from Cas9-expressing transgenic mice. These conditionally immortalized macrophages (CIMs) retain characteristics of primary macrophages derived from the bone marrow yet allow for easy genetic manipulation and a virtually unlimited supply of cells. We demonstrate the utility of this system for dissection of host genetics during intracellular bacterial infection using two important human pathogens: Listeria monocytogenes and Mycobacterium tuberculosis. |
format | Online Article Text |
id | pubmed-6579556 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-65795562019-06-19 Cas9(+) conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond Roberts, Allison W Popov, Lauren M Mitchell, Gabriel Ching, Krystal L Licht, Daniel J Golovkine, Guillaume Barton, Gregory M Cox, Jeffery S eLife Immunology and Inflammation Macrophages play critical roles in immunity, development, tissue repair, and cancer, but studies of their function have been hampered by poorly-differentiated tumor cell lines and genetically-intractable primary cells. Here we report a facile system for genome editing in non-transformed macrophages by differentiating ER-Hoxb8 myeloid progenitors from Cas9-expressing transgenic mice. These conditionally immortalized macrophages (CIMs) retain characteristics of primary macrophages derived from the bone marrow yet allow for easy genetic manipulation and a virtually unlimited supply of cells. We demonstrate the utility of this system for dissection of host genetics during intracellular bacterial infection using two important human pathogens: Listeria monocytogenes and Mycobacterium tuberculosis. eLife Sciences Publications, Ltd 2019-06-17 /pmc/articles/PMC6579556/ /pubmed/31204998 http://dx.doi.org/10.7554/eLife.45957 Text en © 2019, Roberts et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Immunology and Inflammation Roberts, Allison W Popov, Lauren M Mitchell, Gabriel Ching, Krystal L Licht, Daniel J Golovkine, Guillaume Barton, Gregory M Cox, Jeffery S Cas9(+) conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond |
title | Cas9(+) conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond |
title_full | Cas9(+) conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond |
title_fullStr | Cas9(+) conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond |
title_full_unstemmed | Cas9(+) conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond |
title_short | Cas9(+) conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond |
title_sort | cas9(+) conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond |
topic | Immunology and Inflammation |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6579556/ https://www.ncbi.nlm.nih.gov/pubmed/31204998 http://dx.doi.org/10.7554/eLife.45957 |
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