Infectious clones

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 7/04 (2006.01) A61K 39/215 (2006.01) A61K 39/225 (2006.01) A61K 39/295 (2006.01) A61P 31/12 (2006.01) C07K 14/08 (2006.01) C12N 1/21 (2006.01) C12N 15/50 (2006.01) C12N 15/70 (2006.01) C12N 15/86 (2006.01)

Patent

CA 2393325

The present invention relates to methods of preparing a DNA comprising steps, wherein (a) a DNA comprising a full length copy of the genomic RNA (gRNA) or an RNA virus; or (b) a DNA comprising one or several fragments of a gRNA of an RNA virus, which fragments code for an RNA dependent RNA polymerase and at least one structural or non-structural protein; or (c) a DNA having a homology of at least 60% to the sequences of (a) or (b); is cloned into a bacterial artificial chromosome (BAC). Additionally, DNAs are provided, which comprise sequences derived from the genomic RNA (gRNA) of a coronavirus which sequences have a homology of at least 60% to the natural sequence of the virus and code for an RNA dependent RNA polymerase and at least one structural or no- structural protein, wherein a fragment of said DNA is capable of being transcribed into RNA which RNA can be assembled to a virion. Further, the use of these nucleic acids for preparation of viral RNA or virions as well as pharmaceutical preparations comprising these DNAs, viral RNAs or virions is disclosed.

Cette invention a trait à des procédés de préparation d'ADN comprenant des étapes dans le cadre desquelles, il est procédé au clonage dans un chromosome bactérien artificiel (BAC), (a), d'un ADN comprenant une copie de longueur totale d'ARN génomique (ARNg) ou de virus à ARN ou, (b), d'un ADN comprenant un ou plusieurs fragments d'ARNg ou de virus à ARN, lesquels fragments codent une ARN polymérase tributaire de l'ARN et au moins une protéine structurelle ou non structurelle ou, (c), d'un ADN ayant une homologie d'au moins 60 % avec les séquences de (a) ou de (b), L'invention porte, en outre, sur des ADN comprenant des séquences dérivées de l'ARN génomique (ARNg) d'un coronavirus, lesquelles séquences ont une homologie d'au moins 60 % avec la séquence naturelle du virus et codent une ARN polymérase tributaire de l'ARN et au moins une protéine structurelle ou non structurelle, un fragment de cet ADN pouvant être transcrit dans un ARN, lequel ARN peut être joint à un virion. L'invention concerne, de plus, l'utilisation qui est faite de ces acides nucléiques aux fins de la préparation d'ARN viral ou de virions, ainsi que des préparations pharmaceutiques renfermant ces ADN, ARN viraux ou virions.

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