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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.

Elife
|June 18, 2019
PubMed

Insights

Researchers developed a new genome editing system for macrophages, enabling genetic studies of these immune cells. This breakthrough aids research into immunity and infectious diseases like tuberculosis.

Area of Science:

  • Immunology
  • Genetics
  • Cell Biology

Background:

  • Macrophages are crucial for immunity, development, tissue repair, and cancer.
  • Studying macrophage function is challenging due to limitations with current cell models.

Purpose of the Study:

  • To develop a facile system for genome editing in non-transformed macrophages.
  • To enable genetic manipulation of macrophages for functional studies.

Main Methods:

  • Utilized ER-Hoxb8 myeloid progenitors from Cas9-expressing transgenic mice.
  • Differentiated these progenitors into conditionally immortalized macrophages (CIMs).
  • Demonstrated genome editing utility in CIMs.

Main Results:

  • Developed a system for genome editing in non-transformed macrophages.
  • CIMs retain primary macrophage characteristics and allow for easy genetic manipulation.
  • Showcased the system's utility in studying host genetics during infection with Listeria monocytogenes and Mycobacterium tuberculosis.

Conclusions:

  • The developed system provides a valuable tool for studying macrophage biology.
  • Facilitates genetic dissection of host responses to intracellular bacterial pathogens.
  • Overcomes limitations of previous macrophage research models.
Keywords:
Listeria monocytogenesgenome editinghost-pathogen interactionsimmunologyinfectious diseaseinflammationmacrophagemicrobiologymousetuberculosis

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