REVERSE GENETICS USING NON-ENDOGENOUS POL I PROMOTERS
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.
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.