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) and/or at least one 5′-untranslated region element (5′-UTR element), wherein the at least one 3′-UTR element and/or the at least one 5′-UTR element prolongs and/or increases protein production from said artificial nucleic acid molecule and wherein the at least one 3′-UTR element and/or the at least one 5′-UTR element is derived from a stable mRNA. The invention further relates to the use of such an artificial nucleic acid molecule in gene therapy and/or genetic vaccination. Furthermore, methods for identifying a 3′-UTR element and/or a 5′-UTR derived from a stable mRNA element are disclosed.
1 . An artificial nucleic acid molecule comprising:
(a) at least one open reading frame (ORF); and
(b) at least one 3′-untranslated region element (3′-UTR element),
that comprises a nucleic acid sequence that is derived from the 3′-UTR of a transcript of a GNAS (guanine nucleotide-binding protein G subunit alpha) gene and wherein the open reading frame is derived from a gene, which is distinct from a gene from which the at least one 3′-UTR element.
2 . (canceled)
3 . The artificial nucleic acid molecule according to claim 1 comprising at least one 5′-UTR element.
4 . The artificial nucleic acid molecule according to claim 3 , wherein each of the at least one open reading frame, the at least one 3′-UTR element and the at least one 5′-UTR element are heterologous to each other.
5 - 7 . (canceled)
8 . The artificial nucleic acid molecule according to claim 1 , wherein the at least one 3′-UTR element and/or the at least one comprises or consists of a nucleic acid sequence which is derived from the 3′ UTR human or mouse GNAS gene.
9 - 11 . (canceled)
12 . The artificial nucleic acid molecule according to claim 3 , wherein:
(i) the at least one 5′-UTR element is derived from a human or a murine gene selected from the group consisting of: housekeeping genes, genes coding for a membrane protein, genes involved in cellular metabolism, genes involved in transcription, translation and replication processes, genes involved in protein modification and genes involved in cell division; or
(ii) the 5′-UTR is not a 5′ TOP UTR.
13 - 24 . (canceled)
25 . The artificial nucleic acid molecule according to claim 1 , wherein the at least one 3′-UTR element comprises or consists of a nucleic acid sequence which has an identity of at least about 80% a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1 to 4 and SEQ ID NO: 116.
26 . The artificial nucleic acid molecule according claim 1 , wherein the at least one 5′-UTR element comprises or consists of a nucleic acid sequence which has an identity of at least about 80%.
27 . (canceled)
28 . The artificial nucleic acid molecule according to claim 1 , wherein the at least one 3′-UTR element and/or the at least one 5′-UTR element exhibits a length of between 3 and about 500 nucleotides.
29 . The artificial nucleic acid molecule according claim 1 further comprising
c. a poly(A) sequence and/or a polyadenylation signal.
30 - 33 . (canceled)
34 . The artificial nucleic acid molecule according to claim 1 , further comprising a 5′-cap structure, a poly(C) sequence, a histone stem-loop, and/or an IRES-motif.
35 . The artificial nucleic acid molecule according to claim 34 , wherein the histone stem-loop comprises a sequence according to SEQ ID NO: 34.
36 . The artificial nucleic acid molecule according to claim 1 , wherein the nucleic acid comprises a promoter.
37 . The artificial nucleic acid molecule according to claim 1 , wherein the nucleic acid comprises a 5′-TOP UTR.
38 - 41 . (canceled)
42 . A vector comprising an artificial nucleic acid molecule according to claim 1 .
43 - 46 . (canceled)
47 . A cell comprising the artificial nucleic acid molecule according to claim 1 .
48 - 49 . (canceled)
50 . A pharmaceutical composition comprising the artificial nucleic acid molecule according to claim 1 .
51 - 53 . (canceled)
54 . A method for treating or preventing a disorder comprising administering the artificial nucleic acid molecule according to claim 1 to a patient in need thereof.
55 . A method of treating or preventing a disorder comprising transfection of a cell with an artificial nucleic acid molecule according to claim 1 .
56 - 80 . (canceled)
81 . The artificial nucleic acid molecule according to claim 5 , wherein the at least one 5′-UTR element is derived from a 5′-TOP UTR.
82 . The artificial nucleic acid molecule according to claim 81 , wherein the at least one 5′-UTR element comprises a sequence of SEQ ID NO: 33.