RNA polymerase I-mediated expression of viral RNA for the rescue of infectious virulent and avirulent Rift Valley fever viruses.
Rift Valley fever virus (RVFV, Bunyaviridae, Phlebovirus) is a mosquito-transmitted arbovirus that causes human and animal diseases in sub-Saharan Africa and was introduced into the Arabian Peninsula in 2000. Here, we describe a method of reverse genetics to recover infectious RVFV from transfected plasmids based on the use of the cellular RNA polymerase I promoter to synthesize viral transcripts. We compared its efficiency with a system using T7 RNA polymerase and found that both are equally efficient for the rescue of RVFV generating titers of approx. 10(7) to 10(8) pfu/ml. We used the RNA polymerase I-based system to rescue both attenuated MP12 and virulent ZH548 strains as well as chimeric MP12-ZH548 viruses, and in addition RVFV expressing reporter proteins.
Billecocq
Agnès
A
Institut Pasteur, Unité de Génétique Moléculaire des Bunyavirus, 25 rue du Dr Roux, 75724 Paris Cedex, France.
Gauliard
Nicolas
N
Le May
Nicolas
N
Elliott
Richard M
RM
Flick
Ramon
R
Bouloy
Michèle
M
D000818:Animals; D002522:Chlorocebus aethiops; D051379:Mice; D012318:RNA Polymerase I / Q000378:metabolism*; D012367:RNA, Viral / Q000096:biosynthesis*; D012295:Rift Valley Fever / Q000821:virology; D012296:Rift Valley fever virus / Q000235:genetics / Q000254:growth & development* / Q000472:pathogenicity; D016019:Survival Analysis; D014709:Vero Cells; D010948:Viral Plaque Assay; D014773:Virology / Q000379:methods*; D014774:Virulence; D014779:Virus Replication
D012367:RNA, Viral; D012318:RNA Polymerase I