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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.
Abstract

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.

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