Mutated glycoprotein of vesicular stomatitis virus
The invention relates to an isolated non-naturally occurring protein comprising the amino acid sequence as set forth in SEQ ID NO: 1, and wherein the amino acid in position 8, 47, 209 and/or 354 is substituted by any amino acid different from the amino acid indicated at that position in said sequence SEQ ID NO: 1.
1. An isolated non-naturally occurring protein comprising an amino acid sequence at least 90% identical to the amino acid sequence set forth in SEQ ID NO: 1 or a homolog thereof, wherein the homolog thereof comprises an amino acid sequence set forth in any one of SEQ ID NOs: 3, 5, 7, 9, 11, and 13,
wherein the protein comprises an amino acid substitution at one or more of positions 47 and 354 of SEQ ID NO: 1, or at one or more position(s) equivalent to positions 47 and 354 of SEQ ID NO: 1 in any one of SEQ ID NOs: 3, 5, 7, 9, 11, and 13, and
wherein the protein retains the ability to induce membrane fusion and does not interact with LDL membrane receptor.
2. The isolated non-naturally occurring protein according to claim 1 , wherein the protein comprises one of the following amino acid sequences:
SEQ ID NOs: 15-17;
SEQ ID NOs: 21-23;
SEQ ID NOs: 27-29;
SEQ ID NOs: 33-35;
SEQ ID NOs: 39-41;
SEQ ID NOs: 45-47; and
SEQ ID NOs: 51-53.
3. The isolated non-naturally occurring protein according to claim 1 , wherein the amino acid at position 47 or at position 354, or at both positions 47 and 354 of SEQ ID NO: 1 or at equivalent position(s) of the homolog thereof is substituted by an amino acid selected from the group consisting of A, G, F and Q.
4. The isolated non-naturally occurring protein according to claim 1 , comprising amino acid substitutions at positions 47 and 354 of SEQ ID NO: 1 or at equivalent position(s) of the homolog thereof.
5. The isolated non-naturally occurring protein according to claim 1 , further comprising an amino acid substitution at position 8 and/or at position 209 of SEQ ID NO:
1 or at equivalent position(s) of the homolog thereof.
6. The isolated non-naturally occurring protein according to claim 1 , further comprising an amino acid substitution at position 8 of SEQ ID NO: 1 or at equivalent position of the homolog thereof substituted by any amino acid except Q or Y.
7. The isolated non-naturally occurring protein according to claim 1 , further comprising an amino acid substitution at position 209 of SEQ ID NO: 1 or at equivalent position of the homolog thereof substituted by any amino acid except H.
8. The isolated non-naturally occurring protein according to claim 1 , wherein said protein comprises any one of the amino acid sequences set forth in SEQ ID NOs: 57-154.
9. The isolated non-naturally occurring protein according to claim 1 , further comprising an insertion of a heterologous peptide at position 1 or between the amino acids at positions 351 and 352 of SEQ ID NO: 1 or at equivalent position(s) of the homologous protein thereof, wherein the heterologous peptide comprises a part of a ligand or a nanobody that binds a membrane protein expressed on a target cell.
10. The isolated non-naturally occurring protein according to claim 9 , wherein the heterologous peptide is at least a part of a ligand that binds a cellular receptor expressed on a target cell.
11. A nucleic acid molecule encoding the isolated non-naturally occurring protein of claim 1 .
12. A recombinant virus expressing the isolated non-naturally occurring protein of claim 1 .
13. A recombinant virus comprising the nucleic acid molecule of claim 11 .
14. A eukaryotic cell containing or expressing the non-naturally occurring protein of claim 1 , or containing the nucleic acid molecule of claim 11 , or infected by the recombinant virus of claim 12 .
15. The isolated non-naturally occurring protein according to claim 9 , wherein the heterologous peptide comprises an anti-HER2 nanobody, an anti-MUC18 nanobody, or an anti-PD1 nanobody.