Cas9+ conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond.
- Author(s): Roberts, Allison W
- Popov, Lauren M
- Mitchell, Gabriel
- Ching, Krystal L
- Licht, Daniel J
- Golovkine, Guillaume
- Barton, Gregory M
- Cox, Jeffery S
- et al.
Published Web Locationhttps://doi.org/10.7554/elife.45957
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.