Nucleic acid molecules and dual-functional peptides having antiviral activity and delivery activity, compositions and methods thereof
Disclosed are delivery and expression systems of multiple antiviral therapeutic molecules. The therapeutic molecules include a novel class of dual-functional peptide and defective interfering genes of a virus. Also disclosed are compositions comprising the therapeutic molecules that are useful for the treatment and prevention of viral infections. Also disclosed herein are the method of making and using a vector that expresses the therapeutic molecules. Therapeutic molecules include cellular components such as RNA, DNA, peptide, proteins or combination thereof.
1 . A vector comprising one or more viral genes wherein each of the viral genes comprises a deletion to form a detective interfering gene (DIG), the vector expresses one or more nucleic acid molecules that interfere with expression of one or more wild-type viral genes that do not comprise the deletion, the DIG consists of one or more defective genes, that express detective interfering RNAs without any full-length viral RNA and do not generate self-replicable reassortants, wherein
the vector further comprises a dual-functional peptide comprising an HIV-1 Tat Peptide (TAT) and a cationic peptide comprising SEQ ID NO: 4 (P1);
the DIG is a defective viral polymerase gene;
the viral polymerase genes are PB2, PB1 and PA;
the DIG comprises defective PB2, PB1 and PA; and
the DIG comprises SEQ ID NO: 38 (DI-PA), SEQ ID NO: 39 (DI-PB1) and SEQ ID NO: 40 (DI-PB2).
2 . The vector of claim 1 , wherein the deletion is an internal deletion.
3 . The vector of claim 1 , wherein the nucleic acid molecule suppresses replication of a wild-type virus when transfected into cells, animals or humans.
4 . The vector of claim 3 , wherein the replication of the wild-type virus treated with the vector is reduced by about 10-20%, about 20-30%, about 30-40%, about 40-50%, about 50-60%, about 60-70%, about 70-80%, about 80-90%, or about 90-100% as compared to a wild-type virus that is not treated with the vector.
5 . The vector of claim 1 , wherein the vector exerts antiviral activity by preventing endosomal acidification.
6 . A vector comprising one or more viral genes wherein each of the viral genes comprises a deletion to form a detective interfering gene (DIG), said vector expresses one or more nucleic acid molecules that interfere with expression of one or more wild-type viral genes that do not comprise the deletion, the DIG consists of one or more defective genes, that express detective interfering RNAs without any full-length viral RNA and do not generate self-replicable reassortants, wherein
the vector further comprises a dual-functional peptide comprising an HIV-1 Tat Peptide (TAT) and a cationic peptide, comprising SEQ ID NO: 4 (P1);
the DIG is a defective viral polymerase gene;
the viral polymerase genes are PB2, PB1 and PA;
the DIG comprises defective PB2, PB1 and PA; and
the DIG comprises SEQ ID NO: 38 (DI-PA), SEQ ID NO: 39 (DI-PB1) and SEQ ID NO: 40 (DI-PB2),
wherein the deletion is about 50-100 base pair, 100-150 base pair, 150-200 base pair, 200-250 base pair, 250-300 base pair, 300-350 base pair, 350-400 base pair, 400-450 base pair, 450-500 base pair, 500-550 base pair, 550-600 base pair, 600-650 base pair, 650-700 base pair, 700-750 base pair, 750-800 base pair, 800-850 base pair, 850-900 base pair, 900-950 base pair, 950-1000 base pair, 1000-1200 base pair, 1200-1500 base pair, 1500-1800 base pair, or 1800-2100 base pair in length.
7 . The vector of claim 6 , wherein the deletion is an internal deletion.
8 . The vector of claim 6 , wherein the nucleic acid molecule suppresses replication of a wild-type virus when transfected into cells, animals or humans.
9 . The vector of claim 8 , wherein the replication of the wild-type virus treated with the vector is reduced by about 10-20%, about 20-30%, about 30-40%, about 40-50%, about 50-60%, about 60-70%, about 70-80%, about 80-90%, or about 90-100% as compared to a wild-type virus that is not treated with the vector.
10 . The vector of claim 6 , wherein the vector exerts antiviral activity by preventing endosomal acidification.