Artificial nucleic acid molecules
The invention relates to an artificial nucleic acid molecule comprising at least one open reading frame and at least one 3′-untranslated region element (3′-UTR element) comprising a nucleic acid sequence which is derived from the 3′-UTR of a FIG4 gene or from a variant of the 3′-UTR of a FIG4 gene. The invention further relates to the use of such an artificial nucleic acid molecule in gene therapy and/or genetic vaccination. Furthermore, the invention relates to the use of a 3′-UTR element comprising a nucleic acid sequence which is derived from the 3′-UTR of a FIG4 gene or from a variant of the 3′-UTR of a FIG4 gene for the stabilization and/or prolongation of protein expression from a nucleic acid sequence comprising such 3′-UTR element.
1. An artificial nucleic acid molecule comprising
a) at least one open reading frame (ORF) encoding a pathogen antigen, tumor antigen or a human therapeutic protein,
b) at least one Sac Domain-Containing Inositol Phosphatase 3 (FIG4) 3′-untranslated region element (3′-UTR element) comprising a nucleic acid sequence from a human FIG4 gene 3′-UTR, and
c) a poly(A) sequence having a length of 40 to 200 adenine nucleotides,
wherein said ORF is heterologous to said 3′-UTR element and wherein said 3′-UTR element is positioned between said ORF and said poly(A) sequence.
2. A cell comprising the artificial nucleic acid molecule according to claim 1 .
3. The cell according to claim 2 , wherein the cell is a mammalian cell.
4. A pharmaceutical composition comprising the artificial nucleic acid molecule according to claim 1 .
5. The pharmaceutical composition according to claim 4 , further comprising one or more pharmaceutically acceptable vehicles, diluents, excipients, or adjuvants.
6. A kit comprising the artificial nucleic acid molecule according to claim 1 .
7. The kit according to claim 6 , further comprising instructions for use, cells, an adjuvant, means for administration of a pharmaceutical composition, a pharmaceutically acceptable carrier and/or a pharmaceutically acceptable solution for dissolution or dilution of the artificial nucleic acid molecule, a vector, cells, or a pharmaceutical composition.
8. The artificial nucleic acid molecule according to claim 1 , further comprising one or more of a 5′-cap structure, a 5′-UTR, a poly(C) sequence, a histone stem-loop, and an internal ribosome entry site.
9. The artificial nucleic acid molecule according to claim 8 , wherein the histone stem-loop comprises a sequence according to SEQ ID NO: 5.
10. The artificial nucleic acid molecule according to claim 8 , wherein the 5′-UTR is a 5′ terminal oligopyrimidine tract UTR.
11. The artificial nucleic acid molecule according to claim 1 , wherein the G/C content of the open reading frame is increased compared to the wild type open reading frame.
12. The artificial nucleic acid molecule according to claim 1 , wherein the open reading frame comprises a codon-optimized region.
13. The artificial nucleic acid molecule according to claim 1 , wherein the artificial nucleic acid is an mRNA molecule.
14. The artificial nucleic acid molecule according to claim 1 , wherein the open reading frame encodes a human therapeutic protein.
15. The artificial nucleic acid molecule according to claim 1 , wherein the open reading frame encodes a pathogen antigen.
16. The artificial nucleic acid molecule according to claim 1 , wherein the open reading frame encodes a tumor antigen.
17. The artificial nucleic acid molecule according to claim 1 , comprising a sequence according to SEQ ID NO:1 or 2.
18. A vector comprising
a) an open reading frame encoding a pathogen antigen, tumor antigen or a human therapeutic protein,
b) at least one Sac Domain-Containing Inositol Phosphatase 3 (FIG4) 3′-untranslated region element (3′-UTR element) comprising a nucleic acid sequence from a human FIG4 gene 3′-UTR, and
c) a poly(A) sequence having a length of 40 to 200 adenine nucleotides,
wherein said ORF is heterologous to said 3′-UTR element and wherein said 3′-UTR element is positioned between said ORF and said poly(A) sequence.
19. A purified artificial mRNA molecule comprising, from 5′ to 3′,
a) at least one open reading frame (ORF) encoding a pathogen antigen, tumor antigen or a human therapeutic protein,
b) at least one Sac Domain-Containing Inositol Phosphatase 3 (FIG4) 3′-untranslated region element (3′-UTR element) comprising a nucleic acid sequence from a human FIG4 gene 3′-UTR, and
c) a poly(A) sequence having a length of 40 to 200 adenine nucleotides,
wherein said ORF is heterologous to said 3′-UTR element.
20. The purified artificial mRNA molecule according to claim 19 , wherein the open reading frame encodes a human therapeutic protein.
21. The purified artificial mRNA molecule according to claim 19 , wherein the open reading frame encodes a pathogen antigen.
22. The purified artificial mRNA molecule according to claim 19 , wherein the open reading frame encodes a tumor antigen.