Multigenome retroviral vector preparations and methods and systems for producing and using same
First Claim
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1. A recombinant multigenome retroviral vector preparation comprising:
- a. a first retroviral particle comprising two copies of a first retroviral vector genome comprising a first sequence of interest;
b. a second retroviral particle comprising two copies of a second retroviral vector genome comprising a second sequence of interest;
c. a third retroviral particle comprising one copy of the first retroviral vector genome comprising a first sequence of interest and one copy of the second retroviral vector genome comprising a second sequence of interest;
d. a fourth retroviral particle comprising two copies of a third retroviral vector genome comprising a third sequence of interest;
e. a fifth retroviral particle comprising one copy of the first retroviral vector genome comprising a first sequence of interest and one copy of the third retroviral vector genome comprising the third sequence of interest; and
f. a sixth retroviral particle comprising one copy of the second retroviral vector genome comprising a second sequence of interest and one copy of the third retroviral vector genome comprising the third sequence of interest;
wherein the first, second and third sequences of interest are different; and
wherein the retroviral vector preparation is pseudotyped with a heterologous envelope glycoprotein.
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Abstract
The present invention provides novel multigenome retroviral vectors, methods and packaging systems for making such retroviral vectors and methods of use.
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Citations
47 Claims
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1. A recombinant multigenome retroviral vector preparation comprising:
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a. a first retroviral particle comprising two copies of a first retroviral vector genome comprising a first sequence of interest; b. a second retroviral particle comprising two copies of a second retroviral vector genome comprising a second sequence of interest; c. a third retroviral particle comprising one copy of the first retroviral vector genome comprising a first sequence of interest and one copy of the second retroviral vector genome comprising a second sequence of interest; d. a fourth retroviral particle comprising two copies of a third retroviral vector genome comprising a third sequence of interest; e. a fifth retroviral particle comprising one copy of the first retroviral vector genome comprising a first sequence of interest and one copy of the third retroviral vector genome comprising the third sequence of interest; and f. a sixth retroviral particle comprising one copy of the second retroviral vector genome comprising a second sequence of interest and one copy of the third retroviral vector genome comprising the third sequence of interest; wherein the first, second and third sequences of interest are different; and wherein the retroviral vector preparation is pseudotyped with a heterologous envelope glycoprotein. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 29, 41, 42, 43, 44, 45, 46)
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13. A retroviral vector packaging system for producing a pseudotyped multigenome retroviral vector preparation, comprising:
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a. a first nucleic acid molecule encoding a viral envelope protein; b. a second nucleic acid molecule encoding gag and pol proteins; c. a third nucleic acid molecule encoding rev; d. a fourth nucleic acid molecule comprising a first retroviral vector genome comprising a first sequence of interest; e. a fifth nucleic acid molecule comprising a second retroviral vector genome comprising a second sequence of interest; f. a sixth nucleic acid molecule comprising a third retroviral vector genome comprising a third sequence of interest; g. optionally a sixth nucleic acid molecule encoding vpx; and h. a packaging cell or cell line. - View Dependent Claims (14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 47)
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30. A multigenome retroviral vector packaging system, comprising:
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a. at least three nucleic acid molecules each comprising a retroviral vector genome comprising a unique sequence of interest; b. one or more nucleic acid molecules encoding the components necessary to generate pseudotyped retroviral vector particles; c. optionally a nucleic acid molecule encoding vpx; and d. a packaging cell line. - View Dependent Claims (31, 32, 33, 34, 35, 36, 37, 38, 39, 40)
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Specification