Sequence specific DNA recombination in eukaryotic cells
AssigneesPeter Dröge, Boehringer Ingelheim Pharma GmbH & Co. KG
InventorsPeter Droge, Barbara Enenkel
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Abstract
The present invention relates to a method of sequence-specific recombination of DNA in eukaryotic cells, comprising the introduction of a first DNA comprising a nucleotide sequence containing at least one recombination sequence into a cell, introducing a second DNA comprising a nucleotide sequence containing at least one further recombination sequence into a cell, and performing the sequence specific recombination by a bacteriophage lambda integrase Int.
Claims30
A method of promoting sequence specific recombination of DNA in a eukaryotic cell comprising:
- (a) providing a eukaryotic cell comprising a DNA comprising a first attB, attP, attL or attR sequence, or a derivative thereof;
- (b) introducing into the cell a DNA comprising a second attB, attP, attL or attR sequence, or a derivative thereof, into a cell, wherein if said first DNA comprises an attB sequence, or a derivative thereof, said second DNA comprises an attB, attL or attR sequence, or a derivative thereof, or wherein if said first DNA comprises an attP sequence, or a derivative thereof, said second DNA comprises an attP, attL or attR sequence, or a derivative thereof, or wherein if said first DNA comprises an attL sequence, or a derivative thereof, said second DNA comprises an attB, attP or attL sequence, or a derivative thereof, or wherein if said first DNA comprises an attR sequence, or a derivative thereof, said second DNA comprises an attB, attP or attR sequence, or a derivative thereof;
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- (c) providing to the cell a bacteriophage lambda integrase Int under conditions that support integrase-mediated sequence specific recombination.
A method of producing a desired polypeptide in a eukaryotic cell comprising:
- (a) providing a eukaryotic cell comprising a first DNA comprising an attB, attP, attL or attR sequence or a derivative thereof;
- (b) introducing a second DNA comprising an attB, attP, attL or attR sequence or a derivative thereof, and a segment encoding the polypeptide of interest into the cell;
- (c) contacting said cell with a bacteriophage lambda integrase Int under conditions supporting sequence-specific integrase-mediated recombination, wherein the second DNA is integrated into the first DNA;
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- (d) cultivating said cell under conditions, wherein the polypeptide of interest is expressed.
A method of producing a polypeptide of interest in a eukaryotic cell, said cell having at least one naturally-occurring recombination sequence which allows sequence-specific recombination mediated by an bacteriophage lambda Int, or a functional mutant thereof, comprising:
- (a) introducing a DNA comprising an attB, attP, attL or attR sequence or a derivative thereof, and a segment encoding the polypeptide of interest into said cell;
- (b) contacting said cell with a bacteriophage lambda integrase Int under conditions supporting sequence-specific recombination by a bacteriophage lambda integrase Int between the recombination sequence naturally-occurring in said cell and the DNA introduced into said cell;
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- (c) cultivating said cell under conditions, wherein the polypeptide of interest is expressed.
Dependencies are inferred from the claim text.