Use of triplex structure DNA in transferring nucleotide sequences
First Claim
1. A recombinant vector comprising a polynucleotide containing a cis-acting central initiation region (cPPT) and a cis-acting termination region (CTS), these regions being of retroviral or retroviral-like origin, said vector further comprising a defined nucleotide sequence (transgene or sequence of interest) and regulatory signals for reverse transcription, expression, and packaging, wherein said regulatory signals are of retroviral or retroviral-like origin.
2 Assignments
0 Petitions
Accused Products
Abstract
A recombinant vector comprising a polynucleotide containing a cis-acting central initiation region (cPPT) and a cis-acting termination region (CTS) is disclosed. These regions are of retroviral or retroviral-like origin. The vector further comprises a defined nucleotide sequence (transgene or sequence of interest) and regulatory signals for reverse transcription, expression, and packaging, wherein the regulatory signals are of retroviral or retroviral-like origin. In some embodiments the vector is incorporated into compositions for therapeutic purposes. In other embodiments the vector is incorporated into immunogenic compositions, The vector is also useful for methods such as ex vivo transfection or ex vivo transduction of non-mitotic differentiated cells.
80 Citations
41 Claims
- 1. A recombinant vector comprising a polynucleotide containing a cis-acting central initiation region (cPPT) and a cis-acting termination region (CTS), these regions being of retroviral or retroviral-like origin, said vector further comprising a defined nucleotide sequence (transgene or sequence of interest) and regulatory signals for reverse transcription, expression, and packaging, wherein said regulatory signals are of retroviral or retroviral-like origin.
- 5. Recombinant retroviral particles comprising a polynucleotide containing a cis-acting central initiation region (cPPT) and a cis-acting termination region (CTS), these regions being of retroviral or retroviral-like origin and being inserted in a functional orientation with regulatory signals for reverse transcription, wherein said regulatory signals are of retroviral or retroviral-like origin.
-
6. Recombinant retroviral vector particles comprising:
-
(a) a gag polypeptide corresponding to nucleoproteins of a lentivirus or to functional polypeptide derivatives (GAG polypeptides);
(b) a pol polypeptide constituted by the RT, PRO, IN proteins of a lentivirus or a functional polypeptide derivative (POL polypeptide);
(c) an envelope polypeptide or functional polypeptide derivatives (ENV polypeptides);
(d) a recombinant nucleotide sequence comprising a defined nucleotide sequence (transgene or a sequence of interest), placed under the control of regulatory signals for transcription and expression;
a sequence containing regulatory signals for reverse transcription, expression, and packaging, wherein the regulatory signals are of retroviral or retroviral-like origin; and
a polynucleotide comprising a cis-acting central initiation region (cPPT) and a cis-acting termination region (CTS), these regions being of retroviral or retroviral-like origin and being inserted in a functional orientation with said regulatory signals of retroviral or retroviral-like origin.
-
-
19. Recombinant vector particles comprising a recombinant nucleotide sequence comprising a defined nucleotide sequence (transgene), placed under the control of regulatory signals for transcription and expression, regulatory signals for reverse transcription, expression and packaging and a polynucleotide containing a cis-acting central initiation region (cPPT) and a cis-acting termination region (CTS).
-
20. Recombinant vector particles comprising a recombinant nucleotide sequence containing a defined nucleotide sequence (transgene or sequence of interest), placed under the control of regulatory signals for transcription and expression, regulatory signals for reverse transcription, expression and packaging of a retrotransposon and a polynucleotide containing a cis-acting central initiation region (cPPT) and a cis-acting termination region (CTS), these regions being derived from a transposon and being inserted in a functional orientation with the retrotransposon regulatory signals.
-
21. Recombinant retroviral-like particles comprising:
-
a) a polynucleotide containing a cis-acting central initiation region (cPPT) and a cis-acting termination region (CTS), these regions being derived from a retrotransposon and inserted in a functional orientation with retrotransposon regulatory signals;
b) a polypeptide corresponding to the nucleoproteins of a retrotransposon or to functional polypeptide derivatives (GAG polypeptides);
c) a pol polypeptide corresponding to RT, PRO, IN proteins of a retrotransposon or a functional polypeptide derivative (POL polypeptide);
d) a viral envelope polypeptide;
e) a recombinant nucleotide sequence comprising a defined nucleotide sequence (transgene or sequence of interest), placed under the control of regulatory signals for transcription and expression, and regulatory signals for reverse transcription, expression and packaging of the retrotransposon.
-
-
30. A method of constructing a recombinant virus, wherein the method comprises inserting a polynucleotide into a recombinant vector, wherein the polynucleotide comprises a cis-acting central initiation region (cPPT) and a cis-acting termination region (CTS), these regions being of retroviral or retroviral-like origin.
-
31. A method of ex vivo nuclear import of a transgene or nucleotide sequence of interest into a eukaryotic cell, wherein the method comprises constructing a vector, wherein the vector comprises a cis-acting central initiation region (cPPT) and a cis-acting termination region (CTS) wherein the CPPT and CTS regions are of retroviral or retroviral-like origin, and importing the transgene or nucleotide sequence of interest into the cell.
-
34. A polynucleotide comprising a nucleotide sequence derived from a retroviral genome and containing about 80 to 120 nucleotides, preferably 90 to 110 nucleotides, said nucleotide sequence being flanked on one side by a cis-acting central initiation nucleotide sequence (cPPT) and on the other side by a cis-acting central termination nucleotide sequence (CTS).
-
38. A method of transfection or transduction of eukaryotic cells with a transgene or a polynucleotide of interest comprising transfecting or transducing a polynucleotide comprising a cis-acting central initiation region (cPPT) and cis-acting termination region (CTS), these regions being of retroviral or retroviral-like origin, into a eukaryotic cell.
Specification