The Use of CRISPR/Cas9 Gene Editing to Confirm Congenic Contaminations in Host-Pathogen Interaction Studies

Area of Science:

• Microbiology
• Immunology
• Genetics

Background:

• Murine models are crucial for studying bacterial infections like Salmonella Typhimurium.
• Macrophage function in controlling intracellular bacterial replication varies significantly between mouse strains (e.g., C57BL/6, BALB/c).
• Congenic mouse strains, combining genetic backgrounds, can alter infection study outcomes.

Purpose of the Study:

• To investigate how genetic background influences Salmonella Typhimurium infection phenotypes in macrophages.
• To evaluate the utility of CRISPR/Cas9-generated macrophage knockout cell lines in dissecting these genetic contributions.
• To determine the impact of background contaminations on primary macrophage phenotypes in congenic mouse models.

Main Methods:

• Utilizing CRISPR/Cas9 gene editing to create macrophage knockout cell lines.
• Comparing phenotypes of primary macrophages from congenic mice with knockout cell lines.
• Conducting Salmonella Typhimurium infection studies with these genetic models.

Main Results:

• CRISPR/Cas9 technology can identify background contaminations affecting congenic mouse macrophage phenotypes.
• Macrophage knockout cell lines serve as valuable tools to complement existing models.
• Concurrent studies using both genetic models are beneficial for excluding side-effects.

Conclusions:

• CRISPR/Cas9 gene editing is effective in clarifying genetic influences on host-pathogen interactions.
• The use of complementary genetic models enhances the reliability of Salmonella infection studies.
• This methodology improves the interpretation of results from congenic mouse models.