Chemical Synthesis of Poliovirus cDNA: Generation of Infectious Virus in the Absence of Natural Template

Abstract

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.

Structural and functional analysis of virus factories purified from Rabbit vesivirus-infected Vero cells

“…The investigation of enzyme reactions relevant to virus replication, such as RNA synthesis initiation and elongation or the role of VPg in genome replication, which have been extensively studied in the poliovirus system using in vitro cell free systems (Cello et al, 2002;Franco et al, 2005;Molla et al, 1991) have been mostly addressed in caliciviruses using recombinant proteins in vitro (López Vázquez et al, 1998;Machín et al, 2001;Marin et al, 2000;Morales et al, 2004), in the absence of other viral and cellular components which might be crucial for the specificity and efficiency of such processes. The isolation of replication complexes from virus-infected cells could provide a valuable tool for studying, not only the macromolecular components involved in such membranous structures but as an experimental system in which various parameters and or components could be accessible to external manipulation.…”

Una enfermedad lejana: la información sobre poliomielitis y síndrome post-polio en la prensa hispanolusa, 1995-2009

“…El subtítulo dejaba aún más claro lo que la noticia pretendía transmitir: "El experimento, con ADN comprado por correo, levanta el espectro del bioterrorismo". El artículo daba cuenta de la publicación en Science Magazine del trabajo del equipo de Eckard Wimmer en la síntesis del virus de la polio a partir de la información genética disponible sobre el mismo y subrayaba los aspectos más peligrosos del logro (Cello, Aniko, Wimmer, 2002). El propio Wimmer comprendía, en un artículo de 12 de octubre de 2002, los motivos de alarma: "El proyecto fue aprobado en 1998, no tenía nada que ver con el 11 de septiembre.…”

Synthetic biology through the prism of scenarios

“…A feat which has not been achieved yet. Synthetic viruses have been reported but they are not independent life forms [8] and even the transplantation of a genome needs a pre-existing 'empty' host and can thus not be considered ab initio synthesis of 'life' [9]. The concept of synthetic biology has a clear analogy to what engineers do with machines: tinkering around with 'simple' building blocks to create complex assemblies with useful functions [10].…”