Enzymes, cells and methods for site specific recombination at asymmetric sites
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Abstract
The present invention relates to enzymes, compositions and methods for catalyzing asymmetric recombination of non-palindromic recombination sites in a cell free system, in isolated cells or in living organisms. The enzymes and methods of the invention are suitable for mediating specific recombinations between DNA sequences comprising specific recombination sites without being limited to strict palindromic symmetry within each recombination site.
12 Citations
101 Claims
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1-71. -71. (canceled)
- 72. An isolated enzyme capable of mediating a site-specific recombination between two predetermined recombination sites, wherein at least one recombination site is an asymmetric recombination site.
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76. A plurality of isolated enzymes capable of mediating site-specific recombination between two predetermined recombination sites, wherein at least one of the recombination sites is an asymmetric recombination site, wherein at least one enzyme is selected from the group consisting of:
- a wild type recombinase, Cre and Flp mutant, and at least one recombination site is an asymmetric recombination site comprising a spacer sequence selected from the group consisting of;
SEQ ID NOS. 1-34. - View Dependent Claims (77, 78, 79, 80, 81)
- a wild type recombinase, Cre and Flp mutant, and at least one recombination site is an asymmetric recombination site comprising a spacer sequence selected from the group consisting of;
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86. A method for treating a disease, comprising:
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a. providing a composition comprising a DNA molecule comprising a nucleotide sequence encoding at least one enzyme, the at least one enzyme is capable of mediating site-specific excision of a gene fragment flanked between two recombination sites, wherein at least one recombination site is an asymmetric recombination site; b. administering the composition to a subject in need thereof;
thereby obtaining site-specific excision of the gene fragment from a predetermined genomic locus.wherein the composition further comprises a carrier operably connected to the isolated DNA molecule, the carrier capable of targeting said isolated DNA molecule to a cell and promoting internalization of said isolated DNA molecule into the cell and said carrier is selected from the group consisting of;
viruses, liposomes, lipid/DNA complexes, micelles, protein/lipid complexes, nanoparticles, and microparticles. - View Dependent Claims (87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101)
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Specification