Chemical synthesis of poliovirus cDNA: generation of infectious virus in the absence of natural template.
Full-length poliovirus complementary DNA (cDNA) was synthesized by assembling oligonucleotides of plus and minus strand polarity. The synthetic poliovirus cDNA was transcribed by RNA polymerase into viral RNA, which translated and replicated in a cell-free extract, resulting in the de novo synthesis of infectious poliovirus. Experiments in tissue culture using neutralizing antibodies and CD155 receptor-specific antibodies and neurovirulence tests in CD155 transgenic mice confirmed that the synthetic virus had biochemical and pathogenic characteristics of poliovirus. Our results show that it is possible to synthesize an infectious agent by in vitro chemical-biochemical means solely by following instructions from a written sequence.
Department of Molecular Genetics and Microbiology, School of Medicine, State University of New York at Stony Brook, Stony Brook, NY 11794-5222, USA.
Jeronimo
J
Cello
Aniko V
AV
Paul
Eckard
E
Wimmer
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002213:Capsid / Q000378:metabolism; D002474:Cell-Free System; D018076:DNA, Complementary / Q000138:chemical synthesis* / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D005260:Female; D016679:Genome, Viral; D006367:HeLa Cells; D006801:Humans; D008297:Male; D008565:Membrane Proteins; D051379:Mice; D008822:Mice, Transgenic; D009500:Neutralization Tests; D011051:Poliomyelitis / Q000821:virology; D017955:Poliovirus / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity / Q000502:physiology; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000138:chemical synthesis* / Q000235:genetics / Q000502:physiology; D011991:Receptors, Virus / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014158:Transcription, Genetic; D010948:Viral Plaque Assay; D014764:Viral Proteins; D014774:Virulence; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D008565:Membrane Proteins; D012367:RNA, Viral; D011991:Receptors, Virus; D014764:Viral Proteins; C066100:poliovirus receptor; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases