Recombinant viruses comprising the membrane-proximal domain of VSV G protein
AssigneeThe University of Tennessee Research Corporation
InventorsMichael A. Whitt, Clinton S. Robison
Life of this patent
Estimating the term…
Abstract
Recombinant viruses, isolated nucleic acids and methods of generating same encoding for a Rhabdoviral G stem polypeptide are disclosed. Methods, compounds and compositions for target cell fusion potentiation mediated by Rhabdoviral G stem polypeptides, and applications of same are provided.
Claims157
A recombinant Vesicular Stomatitis Virus (VSV) comprising a nucleic acid sequence of a Vesicular Stomatitis Virus (VSV) genome wherein said Vesicular Stomatitis Virus (VSV) genome comprises:
- (a) a deletion in a nucleic acid sequence encoding a Vesicular Stomatitis Virus (VSV) G protein;
- and
- (b) an insertion of an isolated nucleic acid sequence encoding the Vesicular Stomatitis Virus (VSV) G stem polypeptide set forth in SEQ ID NO:
- 2, (ii) an isolated nucleic acid sequence encoding a functional fragment of the Vesicular Stomatitis Virus (VSV) G stem polypeptide set forth in SEQ ID NO:
- 2, (iii) an isolated nucleic acid sequence encoding the amino acid sequence of residues 10-42 of SEQ ID NO:
- 2, or (iv) an isolated nucleic acid sequence that is at least 95% identical to SEQ ID NO:
- 1,
- wherein said deletion comprises the nucleic acid sequence encoding an amino acid sequence of residues 1 to at least 404 of a mature Vesicular Stomatitis Virus (VSV) G protein.
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
A method of producing a recombinant Vesicular Stomatitis Virus (VSV), said recombinant Vesicular Stomatitis Virus (VSV) comprising a deletion in the nucleic acid sequence encoding a Vesicular Stomatitis Virus (VSV) G protein, an insertion of a nucleic acid sequence encoding a Vesicular Stomatitis Virus (VSV) G stem polypeptide fragment and a nucleic acid sequence encoding a heterologous fusion facilitating polypeptide, comprising the steps of:
- (a) contacting a cell with a minivirus comprising cis-acting elements facilitating Vesicular Stomatitis Virus (VSV) replication;
- (b) inserting into said cell a polynucleotide sequence encoding Vesicular Stomatitis Virus (VSV) proteins, a nucleic acid sequence encoding a fragment of a Vesicular Stomatitis Virus (VSV) G stem polypeptide and a nucleic acid sequence encoding a heterologous fusion facilitating polypeptide, wherein said G stem polypeptide fragment potentiates membrane fusion;
- (c) culturing said cell under conditions permitting expression of said nucleic acid sequences to produce said recombinant Vesicular Stomatitis Virus (VSV);
- and
- (d) isolating said recombinant Vesicular Stomatitis Virus (VSV),
- thereby producing said recombinant Vesicular Stomatitis Virus (VSV).
A recombinant virus comprising an isolated nucleic acid sequence encoding
- (a) the Vesicular Stomatitis Virus (VSV) G stem polypeptide set forth in SEQ ID NO:
- 2, an amino acid sequence that is 95% identical to SEQ ID NO:
- 2, a functional fragment of SEQ ID NO:
- 2, or an amino acid sequence of residues 10-42 of SEQ ID NO:
- 2,
- (b) further comprising a nucleic acid sequence encoding a viral fusion facilitating polypeptide,
- wherein said recombinant virus does not comprise a nucleic acid sequence encoding aa 1-404 of a VSV G protein.
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
An isolated nucleic acid molecule encoding the Vesicular Stomatitis Virus (VSV) G stem polypeptide set forth in SEQ ID NO:
- 2, a functional fragment SEQ ID NO:
- 2, an amino acid sequence of residues 10-42 of SEQ ID NO:
- 2, or an amino acid sequence that is at least 95% identical to SEQ ID NO:
- 2.
(canceled)
(canceled)
A method for targeted cell or tissue lysis, comprising the steps of contacting said cell or tissue with an effective amount of a recombinant Vesicular Stomatitis Virus (VSV), wherein a genome of said recombinant Vesicular Stomatitis Virus (VSV) comprises:
- (a) a deletion in the nucleic acid sequence encoding a G protein;
- (b) an insertion of a nucleic acid sequence encoding a Vesicular Stomatitis Virus (VSV) G stem polypeptide fragment, wherein said fragment potentiates membrane fusion;
- (c) a nucleic acid sequence encoding a heterologous fusion facilitating polypeptide;
- and
- (d) a nucleic acid sequence encoding an anti-receptor protein or a polypeptide fragment thereof,
- wherein said anti-receptor protein or polypeptide fragment thereof binds its cognate ligand or receptor, expressed on a surface of said targeted cell or tissue, facilitating targeting of said recombinant Vesicular Stomatitis Virus (VSV) for entry into said cell or tissue, thereby causing targeted cell or tissue lysis.
An isolated nucleic acid sequence encoding the Vesicular Stomatitis Virus (VSV) G stem polypeptide set forth in SEQ ID NO:
- 2 or a functional fragment thereof, wherein said G stem polypeptide or said fragment thereof potentiates membrane fusion.
(canceled)
(canceled)
A recombinant Vesicular Stomatitis Virus (VSV) comprising a nucleic acid sequence of a Vesicular Stomatitis Virus (VSV) genome wherein said Vesicular Stomatitis Virus (VSV) genome comprises:
- (a) a deletion in a nucleic acid sequence encoding a Vesicular Stomatitis Virus (VSV) G protein;
- and
- (b) an insertion of an isolated nucleic acid sequence encoding the Vesicular Stomatitis Virus (VSV) G stem polypeptide set forth in SEQ NO:
- 9, a functional fragment SEQ ID NO:
- 9, or an amino acid sequence that is at least 95% identical to SEQ ID No:
- 9.
(canceled)
(canceled)
(canceled)
A recombinant virus comprising an isolated nucleic acid sequence encoding a Rhabdoviral G stem polypeptide, or a functional fragment thereof, wherein said isolated nucleic acid sequence encodes an amino acid sequence that is at least 95% identical to SEQ ID No:
- 9.
(canceled)
(canceled)
An isolated nucleic acid molecule comprising a nucleic acid sequence encoding a Vesicular Stomatitis Virus (VSV) G stem polypeptide or a fragment thereof, wherein said Vesicular Stomatitis Virus (VSV) G stem polypeptide or fragment thereof is at least 95% homologous to SEQ ID No:
- 9, and a nucleic acid sequence encoding a heterologous fusion facilitating polypeptide.
(canceled)
(canceled)
(canceled)
Dependencies are inferred from the claim text.