Generation of viral transfectants using recombinant...

C - Chemistry – Metallurgy – 12 – N

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Details

C12N 15/86 (2006.01) C07K 14/11 (2006.01) C12N 5/10 (2006.01) C12N 7/00 (2006.01) C12N 7/01 (2006.01) C12N 7/04 (2006.01) C12N 9/12 (2006.01) C12N 15/87 (2006.01) A61K 39/00 (2006.01) A61K 48/00 (2006.01)

Patent

CA 2253595

This invention provides a method of producing viral transfectants (viruses containing a heterologous nucleic acid) that eliminates the need for purified RNP complexes or for purified viral RNA polymerases. The method generally involves: (i) combining a ribonucleic acid (nucleic acid transcript) with an isolated nucleoprotein (NP) of the virus to form a synthetic ribonucleoprotein complex (RNP); (ii) transfecting a host cell with the synthetic ribonucleoprotein complex; and (iii) holding (culturing) the host cell under conditions permitting replication of the virus.

Cette invention se rapporte à un procédé pour produire des transfectants viraux (virus contenant un acide nucléique hétérologue) qui permet d'éliminer la nécessité de recourir à des complexes de ribonucléoprotéines (RNP) purifiés ou à des ARN-polymérases virales purifiées. Ce procédé, d'une façon générale, consiste: (i) à combiner un acide ribonucléique (acide nucléique de transcription) avec une nucléoprotéine (NP) isolée du virus, afin de former un complexe de ribonucléoprotéines (RNA) synthétique; (ii) à effectuer la transfection d'une cellule hôte avec ce complexe de ribonuncléoprotéines synthétique; et (iii) à maintenir (cultiver) cette cellule hôte dans des conditions permettant la réplication dudit virus.

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