US2016024525A1PendingUtilityA1

Reverse genetics using non-endogenous pol i promoters

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Assignee: NOVARTIS AGPriority: May 21, 2009Filed: May 7, 2015Published: Jan 28, 2016
Est. expiryMay 21, 2029(~2.8 yrs left)· nominal 20-yr term from priority
Inventors:Philip DormitzerMichael FrantiPirada SuphaphiphatPeter MasonBjoern KeinerStefania Crotta
A61P 31/14A61P 31/16A61P 37/04A61K 39/12C12N 2760/16134C12N 2760/16021A61K 39/145C12N 2760/16034C12N 2830/85C12N 2830/00C12N 7/00C12N 2760/16234C12N 15/85C12N 2760/16043C12N 15/86C12N 2760/16052C12N 5/12
43
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Claims

Abstract

Expression of a transgene is driven in a host cell using a pol I promoter which is not endogenous to an organism from the same taxonomic order from which the host cell is derived.

Claims

exact text as granted — not AI-modified
1 : A host cell comprising at least one expression construct encoding a viral RNA molecule, wherein expression of the viral RNA molecule from the construct is controlled by a pol I promoter which is not endogenous to the host cell's taxonomic order. 
     
     
         2 : A cell having at least one endogenous pol I promoter which control(s) expression of endogenous rRNA and at least one non-endogenous pol I promoter which control(s) expression of a viral RNA or the complement thereof. 
     
     
         3 : A method for producing a recombinant virus, comprising a step of growing the cell of  claim 1  under conditions where the viral RNA molecule is expressed in order to produce virus. 
     
     
         4 : The method of  claim 3  further comprising infecting a culture host with the virus produced by growing the cell; culturing the culture host infected with the virus to produce further virus; and purifying the further virus obtained by culturing the infected culture host. 
     
     
         5 : The method of  claim 4  further comprising preparing a vaccine from the purified further virus. 
     
     
         6 : The cell of  claim 1 , wherein the pol I promoter is a primate pol I promoter and the cell is a non-primate cell. 
     
     
         7 : The cell of  claim 1 , wherein the pol I promoter is a non-canine pol I promoter and the cell is a canine cell. 
     
     
         8 : The cell of  claim 7  wherein the pol I promoter is a human pol I promoter. 
     
     
         9 : The cell of  claim 8  wherein the cell is an MDCK cell. 
     
     
         10 : The cell of  claim 9  wherein the MDCK cell is cell line MDCK 33016 (DSM ACC2219). 
     
     
         11 : The cell of  claim 1 , wherein the cell includes at least one bi-directional expression construct. 
     
     
         12 : The cell of  claim 1 , wherein the expression construct is an expression vector or a linear expression construct. 
     
     
         13 : The cell of  claim 1 , wherein the virus is a segmented virus. 
     
     
         14 : The cell of  claim 1 , wherein the virus is a non-segmented virus. 
     
     
         15 : The cell of  claim 1 , wherein the virus is a negative-strand RNA virus. 
     
     
         16 : The cell of  claim 15 , wherein the virus is influenza virus.

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