Nucleic acid comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal for increasing the expression of an encoded pathogenic antigen
The present invention relates to a nucleic acid sequence, comprising or coding for a coding region, encoding at least one peptide or protein comprising a pathogenic antigen or a fragment, variant or derivative thereof, at least one histone stem-loop and a poly(A) sequence or a polyadenylation signal. Furthermore the present invention provides the use of the nucleic acid for increasing the expression of said encoded peptide or protein. It also discloses its use for the preparation of a pharmaceutical composition, especially a vaccine, e.g. for use in the treatment of infectious diseases. The present invention further describes a method for increasing the expression of a peptide or protein comprising a pathogenic antigen or a fragment, variant or derivative thereof, using the nucleic acid comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal.
1. A composition comprising a mRNA encoding a Severe Acute Respiratory Syndrome (SARS) coronavirus spike (S) protein coding sequence and a histone stem loop sequence, said mRNA provided in complex with a lipid.
2. The composition of claim 1 , wherein the mRNA comprises a Poly-A sequence.
3. The composition of claim 2 , wherein the Poly-A sequence is about 25 to 400 adenosine nucleotides.
4. The composition of claim 1 , wherein the mRNA comprises a 5′ untranslated region (UTR) and/or a 3′ UTR.
5. The composition of claim 4 , wherein the mRNA comprises a 5′ untranslated region (UTR) and a 3′ UTR.
6. The composition of claim 1 , wherein the lipid comprises a cationic lipid.
7. The composition of claim 1 , wherein the coding sequence of the mRNA has a G/C content that is increased compared with the G/C content of the coding sequence of the wild-type RNA.
8. The composition of claim 1 , wherein the mRNA comprises a nucleotide analog substitution.
9. The composition of claim 1 , wherein the mRNA comprises a methylated 5′ cap.
10. The composition of claim 8 , wherein the nucleotide analog substitution comprises at least one pseudouridine-5′-triphosphate substitution.
11. The composition of claim 1 , further comprising a polyethyleneglycol.
12. The composition of claim 1 , further comprising an adjuvant.
13. The composition of claim 1 , wherein the mRNA comprises a methylated 5′ cap; a 5′ UTR; a 3′ UTR and a Poly-A sequence of about 25 to 400 adenosine nucleotides.
14. The composition of claim 13 , wherein the mRNA has a G/C content that is increased compared with the G/C content of the coding sequence of the wild-type RNA.
15. The composition of claim 14 , wherein the lipid comprises a cationic lipid.
16. The composition of claim 1 , wherein the histone stem loop sequence is at least 90% identical to the sequence of SEQ ID NO: 39 or SEQ ID NO: 42.
17. The composition of claim 16 , wherein the histone stem loop sequence is at least 90% identical to the sequence of SEQ ID NO: 39.
18. A composition comprising a purified mRNA encoding: a 5′ UTR; a Severe Acute Respiratory Syndrome (SARS) coronavirus spike (S) protein coding sequence; a 3′ UTR; a Poly-A sequence of 25 to 400 adenosine nucleotides; and a histone stem loop sequence, said mRNA comprising a 5′ Cap, wherein the purified mRNA is provided in complex with a cationic lipid.
19. The composition of claim 18 , wherein the mRNA comprises a methylated 5′ cap.
20. The composition of claim 18 , wherein the mRNA has a G/C content that is increased compared with the G/C content of the coding sequence of the wild-type RNA.
21. The composition of claim 18 , wherein the histone stem loop sequence is at least 90% identical to the sequence of SEQ ID NO: 39.