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Updated: Jan 23, 2026

Delivery of the Cas9/sgRNA Ribonucleoprotein Complex in Immortalized and Primary Cells via Virus-like Particles "Nanoblades"
Published on: March 31, 2021
Cas9+ conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond
Allison W Roberts1, Lauren M Popov1, Gabriel Mitchell1
1Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States.
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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