| CPC C07K 14/005 (2013.01) [C12N 7/00 (2013.01); C12N 15/86 (2013.01); A61K 38/00 (2013.01); C12N 2730/10122 (2013.01); C12N 2730/10123 (2013.01); C12N 2730/10133 (2013.01); C12N 2750/14143 (2013.01)] | 18 Claims |
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1. A recombinant nucleic acid molecule comprising:
a negative strand nucleic acid molecule or a pregenomic RNA (pgRNA) nucleic acid molecule encoding a chemokine, a cytokine, an apoptosis inducing protein, a diphtheria toxin A, a diphtheria toxin A fragment, or any combination thereof, flanked by a first viral transcription recognition signal and a second viral transcription recognition signal, wherein the negative strand nucleic acid molecule or pgRNA nucleic acid molecule is from a virus selected from the group consisting of hepatitis B virus, influenza virus and coronavirus;
a first promoter upstream (5′) of the first viral transcription recognition signal and the 3′ end of the negative strand nucleic acid molecule or pgRNA nucleic acid molecule, and
a second promoter located between the second viral transcription recognition signal and the 5′ end of the negative strand nucleic acid molecule or pgRNA nucleic acid molecule encoding a chemokine, a cytokine, an apoptosis inducing protein, a diphtheria toxin A, a diphtheria toxin A fragment, or any combination thereof,
wherein
(i) the first promoter and the second promoter are oriented in opposite direction and toward each other;
(ii) the second promoter and the second viral transcription recognition signal are upstream (5′) of the 5′ end of the negative strand nucleic acid molecule or pgRNA nucleic acid molecule;
(iii) the first and second viral transcription recognition signals are oriented in the same direction; and
(iv) wherein
a. if the negative strand nucleic acid molecule or pgRNA nucleic acid molecule is from a hepatitis B virus, each of the first and second viral transcription recognition signals comprises the epsilon recognition signal sequence of SEQ ID NO: 1;
b. if the negative strand nucleic acid molecule or pgRNA nucleic acid molecule is from an influenza virus, the second viral transcription recognition signal comprises the sequence of SEQ ID NO: 22 and the first viral transcription recognition signal comprises the sequence of SEQ ID NO: 23; or
c. if the negative strand nucleic acid molecule or pgRNA nucleic acid molecule is from a coronavirus, each of the first and second viral transcription recognition signals comprises the sequence of SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 28, or SEQ ID NO: 30.
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