Approaches for timeline reductions in pathogenesis studies using genetically modified mice

Area of Science:

• Microbiology
• Genetics
• Immunology

Background:

• Genetically modified mouse models are crucial for microbiology research but are costly and time-consuming to generate.
• While in vivo clustered regularly interspaced short palindromic repeats (CRISPR) technology promises faster genome manipulation, its impact on experimental timelines remained unclear.

Purpose of the Study:

• To optimize an in oviduct murine genetic manipulation technique for use by microbiologists.
• To establish a rapid protocol for generating knockout mice and testing host factors in disease models.

Main Methods:

• Optimization of an in oviduct murine genetic manipulation technique.
• Generation of knockout mice using this optimized protocol.
• Utilizing an influenza A virus infection model to assess host factor importance.

Main Results:

• A protocol was detailed for generating knockout mice and testing host factors in as little as 11 weeks.
• The method allows for preliminary host factor importance testing before a fully backcrossed knockout line is established (~22 weeks).

Conclusions:

• This optimized in vivo CRISPR-based approach significantly reduces the time and resources needed for generating genetically modified mice in microbiology.
• Wider adoption will accelerate the discovery of pathogenic mechanisms and the development of novel therapies for infectious diseases.