C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/86 (2006.01) C12N 5/10 (2006.01) C12N 7/01 (2006.01) C12N 9/22 (2006.01) C12N 15/861 (2006.01)
Patent
CA 2375098
A method of producing recombinant viruses which require helper viruses for packaging is provided. The method uses a recombinant helper virus which has been designed to contain at least one rare-cutting restriction site (e.g., for I-SceI), a viral vector which is helper-dependent for packaging, and a recombinant host cell which is capable of expressing a rare-cutting restriction enzyme (e.g., I-SceI). The method involves transfecting or infecting the host cell with the helper virus and viral vector and incubating the cell under conditions which permit encapsidation of the viral vector in an adenovirus capside. Thereafter, the I-SceI expressed by the host cell digests the helper virus, permitting ready separation of the digested fragments of the helper virus from the packaged recombinant virus.
L'invention concerne une méthode de fabrication de virus recombinés nécessitant des virus auxiliaires pour l'encapsidation. La méthode fait intervenir un virus auxiliaire recombiné qui a été conçu pour pouvoir contenir au moins un site de restriction rare (par exemple, I-SceI), un vecteur viral qui dépend de l'auxiliaire pour l'encapsidation, et une cellule hôte recombinée pouvant exprimer un enzyme de restriction rare (par exemple, I-SceI). La méthode consiste à transfecter ou à infecter la cellule hôte avec le virus auxiliaire et le vecteur viral et à incuber la cellule dans des conditions qui permettent l'encapsidation du vecteur viral dans une capside d'adénovirus. Ensuite, le I-SceI exprimé par la cellule hôte digère le virus auxiliaire, ce qui permet une séparation immédiate des fragments digérés du virus auxiliaire d'avec le virus recombiné.
Wilson James M.
Xiao Weidong
Gowling Lafleur Henderson Llp
Trustees Of The University Of Pennsylvania
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