Related Experiment Videos
A Human DPP4-Knockin Mouse's Susceptibility to Infection by Authentic and Pseudotyped MERS-CoV
Changfa Fan1, Xi Wu2, Qiang Liu3
1Division of Animal Model Research, Institute for Laboratory Animal Resources, National Institutes for Food and Drug Control, Beijing 100050, China. fancf@nifdc.org.cn.
Abstract:
Infection by the Middle East respiratory syndrome coronavirus (MERS-CoV) causes respiratory illness and has a high mortality rate (~35%). The requirement for the virus to be manipulated in a biosafety level three (BSL-3) facility has impeded development of urgently-needed antiviral agents. Here, we established anovel mouse model by inserting human dipeptidyl peptidase 4 (hDPP4) into the Rosa26 locus using CRISPR/Cas9, resulting in global expression of the transgene in a genetically stable mouse line. The mice were highly susceptible to infection by MERS-CoV clinical strain hCoV-EMC, which induced severe diffuse pulmonary disease in the animals, and could also be infected by an optimized pseudotyped MERS-CoV. Administration of the neutralizing monoclonal antibodies, H111-1 and m336, as well as a fusion inhibitor peptide, HR2P-M2, protected mice from challenge with authentic and pseudotyped MERS-CoV. These results confirmed that the hDPP4-knockin mouse is a novel model for studies of MERS-CoV pathogenesis and anti-MERS-CoV antiviral agents in BSL-3 and BSL-2facilities, respectively.
Insights
A new mouse model expressing human DPP4 allows MERS-CoV research outside high-security labs. This breakthrough facilitates the development of effective treatments for Middle East respiratory syndrome coronavirus (MERS-CoV) infections.
Area of Science:
- Virology
- Immunology
- Genetics
Background:
- Middle East respiratory syndrome coronavirus (MERS-CoV) infection presents a significant public health threat with high mortality.
- Research into MERS-CoV antivirals is hindered by the need for Biosafety Level 3 (BSL-3) facilities.
Purpose of the Study:
- To develop a novel mouse model for studying MERS-CoV pathogenesis and antiviral therapies.
- To enable MERS-CoV research in lower biosafety level facilities.
Main Methods:
- CRISPR/Cas9 gene editing was used to knock in human dipeptidyl peptidase 4 (hDPP4) into the Rosa26 locus of mice.
- The resulting genetically stable hDPP4-knockin mice were challenged with MERS-CoV clinical strains and pseudotyped MERS-CoV.
- Therapeutic efficacy of neutralizing monoclonal antibodies (H111-1, m336) and a fusion inhibitor peptide (HR2P-M2) was evaluated.
Main Results:
- The hDPP4-knockin mice exhibited high susceptibility to MERS-CoV infection, developing severe diffuse pulmonary disease.
- Infection was also established using an optimized pseudotyped MERS-CoV, enabling studies in Biosafety Level 2 (BSL-2) conditions.
- Administration of monoclonal antibodies and the fusion inhibitor peptide conferred significant protection against MERS-CoV challenge.
Conclusions:
- The hDPP4-knockin mouse represents a valuable new model for MERS-CoV research.
- This model facilitates the study of MERS-CoV pathogenesis and the evaluation of antiviral agents in BSL-3 and BSL-2 settings.
- The developed model accelerates the discovery and development of urgently needed MERS-CoV therapeutics.