Reverse Genetics for Crimean-Congo Hemorrhagic Fever Virus

Author:

Flick Ramon123,Flick Kirsten1,Feldmann Heinz12,Elgh Fredrik345

Affiliation:

1. Special Pathogens Program, National Microbiology Laboratory, Health Canada, Canadian Science Centre for Human and Animal Health, Winnipeg, Canada R3E 3R2

2. Department of Medical Microbiology, University of Manitoba, Winnipeg, Canada R3E 0W3

3. Center for Microbiological Preparedness, Swedish Institute for Infectious Disease Control, SE-171 82 Solna

4. Swedish Defence Research Agency, SE-901 82 Umeå

5. Department of Medical Biosciences/Pathology, Umeå University, SE-901 85 Umeå, Sweden

Abstract

ABSTRACT The widespread geographical distribution of Crimean-Congo hemorrhagic fever (CCHF) virus (more than 30 countries) and its ability to produce severe human disease with high mortality rates (up to 60%) make CCHF a major public health concern worldwide. We describe here the successful establishment of a reverse genetics technology for CCHF virus, a member of the genus Nairovirus , family Bunyaviridae . The RNA polymerase I (pol I) system was used to generate artificial viral RNA genome segments (minigenomes), which contained different reporter genes in antisense (virus RNA) or sense (virus-complementary RNA) orientation flanked by the noncoding regions of the CCHF virus S segment. Reporter gene expression was observed in different eukaryotic cell lines following transfection and subsequent superinfection with CCHF virus, confirming encapsidation, transcription, and replication of the pol I-derived minigenomes. The successful transfer of reporter gene activity to fresh cells demonstrated the generation of recombinant CCHF viruses, thereby confirming the packaging of the pol I-derived minigenomes into progeny viruses. The system offers a unique opportunity to study the biology of nairoviruses and to develop therapeutic and prophylactic measures against CCHF infections. In addition, we demonstrated for the first time that the human pol I system can be used to develop reverse genetics approaches for viruses in the family Bunyaviridae . This is important since it might facilitate the manipulation of bunyaviruses with cell and host tropisms restricted to primates.

Publisher

American Society for Microbiology

Subject

Virology,Insect Science,Immunology,Microbiology

Reference46 articles.

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