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US 12,398,404 B2Granted
Methods and constructs for transient production of lentiviral vector
AssigneeLonza Walkersville, Inc.
InventorsYoung Shin, Anandita Seth
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Abstract
The present disclosure relates to methods for producing lentiviral vectors using mammalian cells. Specifically, the methods utilize three plasmids, rather than four, to provide the required packaging elements and transfer vector to a cell, allowing for the production of a large number of lentiviral vectors in mammalian cells, including suspension-based cells. These methods allow for the production of lentiviral vectors that can be tailored to include a specific gene of interest.
Claims20
A method of producing a lentiviral vector, comprising:
- a. transfecting a mammalian cell with:
- i. a first nucleic acid molecule encoding a lentiviral regulator of expression of virion proteins (REV) gene under control of a first promoter and an envelope glycoprotein gene under control of a second promoter;
- ii. a second nucleic acid molecule encoding a gene of interest under control of a third promoter;
- and
- iii. a third nucleic acid molecule encoding a lentiviral group specific antigen (GAG) gene and a lentiviral polymerase (POL) gene both under control of a fourth promoter,
- wherein the first, second, and third nucleic acid molecules are three separate nucleic acid molecules;
- b. culturing the transfected mammalian cell;
- and
- c. isolating the lentiviral vector.
6 more elements
Dependencies are inferred from the claim text.