Intergenic regions as insertion sites in the genome of modified vaccinia virus ankara (MVA)
The present invention relates to novel insertion sites useful for the integration of exogenous sequences into the Modified Vaccinia Ankara (MVA) virus genome. The present invention further provides plasmid vectors to insert exogenous DNA into the genome of MVA. Furthermore, the present invention provides recombinant MVA comprising an exogenous DNA sequence inserted into the new insertion site as medicine or vaccine.
1. A recombinant Modified Vaccinia Ankara (MVA) virus comprising a heterologous DNA sequence inserted into an intergenic region (IGR) of the viral genome, wherein the IGR is IGR 088R-089L.
2. The recombinant MVA virus of claim 1 , wherein the heterologous DNA sequence comprises a coding sequence.
3. The recombinant MVA virus of claim 2 , wherein the coding sequence is placed under the transcriptional control of a poxviral transcription control element.
4. The recombinant MVA virus of claim 2 , wherein the coding sequence encodes at least one protein, polypeptide, peptide, foreign antigen, or antigenic epitope.
5. The recombinant MVA virus of claim 1 , wherein the heterologous DNA sequence comprises a sequence from Dengue virus.
6. The recombinant MVA virus of claim 1 , wherein the heterologous DNA sequence comprises a sequence from Japanese encephalitis virus.
7. The recombinant MVA virus of claim 1 , wherein the heterologous DNA sequence comprises a sequence from Hepatitis virus B.
8. The recombinant MVA virus of claim 1 , wherein the heterologous DNA sequence comprises a sequence from Hepatitis virus C.
9. The recombinant MVA virus of claim 1 , wherein the heterologous DNA sequence comprises a sequence from human immunodeficiency virus (HIV).
10. The recombinant MVA virus of claim 5 , wherein the Dengue virus sequence is selected from the group consisting of NS1 and PrM sequences.
11. An immunogenic composition comprising the recombinant MVA virus of claim 1 and a physiologically acceptable carrier, diluent, adjuvant, or additive.
12. The immunogenic composition of claim 11 , wherein the heterologous DNA sequence comprises a sequence from Dengue virus, Japanese encephalitis virus, Hepatitis virus B, Hepatitis virus C, or human immunodeficiency virus (HIV).
13. An isolated cell comprising the recombinant MVA virus of claim 1 .
14. A plasmid vector comprising a DNA sequence derived from or homologous to the genome of an MVA virus, wherein the DNA sequence comprises a complete IGR or a fragment of an IGR, wherein inserted into said IGR is a cloning site for the insertion of an exogenous DNA sequence, and wherein the IGR is IGR 088R-089L.
15. An isolated cell comprising the plasmid vector of claim 14 .
16. A method for inducing an immune response in an animal comprising administering the recombinant modified vaccinia Ankara virus of claim 1 to an animal.
17. The method of claim 16 , wherein the heterologous DNA sequence comprises a sequence from Dengue virus, Japanese encephalitis virus, Hepatitis virus B, Hepatitis virus C, or human immunodeficiency virus (HIV).
18. A method for introducing a heterologous DNA sequence into a cell comprising infecting the cell with the recombinant modified vaccinia Ankara virus of claim 1 .
19. The method of claim 18 , wherein the heterologous DNA sequence comprises a sequence from Dengue virus, Japanese encephalitis virus, Hepatitis virus B, Hepatitis virus C, or human immunodeficiency virus (HIV).
20. A method for producing a recombinant modified vaccinia Ankara virus comprising:
(a) transfecting a cell with the plasmid vector of claim 14 ;
(b) infecting the transfected cell with an MVA; and
(c) isolating a recombinant MVA comprising the heterologous DNA.