DNA recombination in eukaryotic cells by the bacteriophage PHIC31 recombination system
First Claim
1. A eukaryotic cell that comprises a prokaryotic recombinase polypeptide or a nucleic acid that encodes a prokaryotic recombinase, wherein the recombinase can mediate site-specific recombination between a first recombination site and a second recombination site that can serve as a substrate for recombination with the first recombination site, but in the absence of an additional factor that is not present in the eukaryotic cell cannot mediate recombination between two hybrid recombinase recombination sites that are formed upon recombination between the first recombination site and the second recombination site.
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Abstract
This invention provides methods for obtaining specific and stable integration of nucleic acids into eukaryotic cells. The invention makes use of site-specific recombination systems that use prokaryotic recombinase polypeptides, such as the ΦC31 integrase, that can mediate recombination between the recombination sites, but not between hybrid recombination sites that are formed upon the recombination. Thus, the recombination is irreversible in the absence of additional factors. Eukaryotic cells that contain the recombinase polypeptides, or genes that encode the recombinases, are also provided.
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Citations
47 Claims
- 1. A eukaryotic cell that comprises a prokaryotic recombinase polypeptide or a nucleic acid that encodes a prokaryotic recombinase, wherein the recombinase can mediate site-specific recombination between a first recombination site and a second recombination site that can serve as a substrate for recombination with the first recombination site, but in the absence of an additional factor that is not present in the eukaryotic cell cannot mediate recombination between two hybrid recombinase recombination sites that are formed upon recombination between the first recombination site and the second recombination site.
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13-31. -31. (canceled).
- 32. A nucleic acid that comprises a polynucleotide sequence that encodes a bacterial recombinase polypeptide operably linked to a promoter that functions in a eukaryotic cell, wherein the recombinase polypeptide cannot mediate recombination between two hybrid recombination sites that are formed upon recombination between a first recombination site and a second recombination site in the absence of an additional factor.
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36. A eukaryotic cell that comprises a polynucleotide that comprises a first bacteriophage Φ
- C31 recombination site.
- View Dependent Claims (37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47)
Specification