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US 2014/0193914 A1Published application
Introduction of Modular Vector Elements During Production of a Lentivirus
AssigneeThermo Fisher Scientific Biosciences Inc.
InventorsJamie Cearley, Rusla DuBreuil
Life of this patent
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Abstract
Methods and kits for lentiviral production are provided. By separating certain components of the vector including but not limited to promoter, reporter and selection marker preference components from the genetic content of interest, one can create libraries that allow for the efficient generation of custom selected lentiviral vectors.
Claims13
A kit for the introduction of modular vector elements comprising:
- (a) a preference vector, wherein said preference vector comprises a promoter sequence;
- and
- (b) a content vector, wherein said content vector comprises a genomic content of interest, a first LTR sequence that is 5′ of the genomic content of interest and a second LTR sequence that is 3′ of the genomic content of interest, and wherein said content vector does not comprise any promoter sequence and said preference vector does not comprise said genomic content of interest and further wherein said preference vector and said content vector each contain a single recombination site and are capable of combining through a cross-over event to form a lentiviral vector.
A kit for the introduction of modular vector elements comprising:
- (a) a plurality of preference vectors, wherein said preference vectors each comprise a promoter sequence and a single recombination site;
- (b) a plurality of cells from a packaging cell line;
- and
- (c) at least one content vector, wherein each content vector comprises a sequence corresponding to a genomic content of interest, a single recombination site, a first LTR sequence that is 5′ of the genomic content of interest and a second LTR sequence that is 3′ of the genomic content of interest, and wherein each content vector has an absence of any promoter sequences, and each preference vector does not comprise any sequence that corresponds to said genomic content of interest, and further, wherein at least two preference vectors have different promoter sequences.
1 more element
A method for generating a viral vector comprising;
- (a) introducing a preference vector, into a cell, wherein said preference vector contains a single recombination site and wherein the cell comprises one or more packaging plasmids;
- (b) introducing a content vector into said cell, wherein said content vector contains a single recombination site, a sequence that corresponds to genomic content of interest, a first LTR sequence 5′ of the genomic content of interest and a second LTR sequence 3′ of the genomic content of interest;
- and
- (c) subjecting said cell to conditions under which the preference vector and the content vector are capable of combining into a viral transcript, wherein said viral transcript is a lentiviral vector.
1 more element
Dependencies are inferred from the claim text.