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Murine Norovirus: Additional Protocols for Basic and Antiviral Studies.

Christiane E Wobus1, Amy M Peiper2, Alice M McSweeney3

  • 1Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan.

Current Protocols
|July 21, 2023
PubMed
Summary
This summary is machine-generated.

Murine norovirus (MNV) research is advanced with new protocols for studying its biology. These methods cover MNV genome quantification, recombinant virus generation, protein expression, and infection models like enteroids and neonatal mice.

Keywords:
baculovirus expressioncircular polymerase extension reactionmurine norovirusneonatal infectionstuft cell-enriched murine intestinal enteroid culture

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Area of Science:

  • Virology
  • Gastroenterology
  • Molecular Biology

Background:

  • Murine norovirus (MNV) is a key model for human noroviruses, responsible for widespread gastroenteritis.
  • Studying MNV is crucial due to technical challenges with human noroviruses.
  • MNV offers a cell culture and reverse genetics system for in-host infection studies.

Purpose of the Study:

  • To present updated and novel techniques for studying murine norovirus (MNV) biology.
  • To expand upon existing protocols for MNV research.
  • To facilitate deeper understanding of norovirus mechanisms.

Main Methods:

  • Indirect assays for cell cytotoxicity and antiviral activity.
  • RT-qPCR for measuring MNV genome titers.
  • Generation of recombinant MNV via minimal passaging and CPER.
  • Expression of MNV NS1-2 protein in insect and mammalian cells.
  • Infection of enteroids and neonatal mice to model MNV-induced diarrhea.

Main Results:

  • Established protocols for quantifying MNV genome and assessing antiviral efficacy.
  • Developed methods for generating infectious recombinant MNV strains.
  • Enabled expression and purification of key MNV proteins (NS1-2).
  • Validated enteroid and neonatal mouse models for MNV infection studies.

Conclusions:

  • The described protocols provide a comprehensive toolkit for advancing MNV research.
  • These techniques enhance the study of MNV replication, pathogenesis, and antiviral strategies.
  • The updated methods support the use of MNV as a model for human norovirus infections.