Methods and compositions for targeted gene modification
First Claim
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1. A method of placing an exogenous nucleic acid into DNA of a cell comprising introducing into the cell:
- (i) an exogenous nucleic acid,(ii) a proteinaceous fusion molecule that comprisesa recombinase domain,a protein-protein interaction domain, anda nuclear localization signal domain, and(iii) a single strand DNA probe complementary to a target site in DNA of the cell (cssDNA), with the probe and the recombinase domain complexed as a nucleoprotein filament at the time of the introduction into the cell,with the exogenous nucleic acid sequence being chosen for incorporation into the DNA of the cell and designed to be free of specific binding to the proteinaceous fusion molecule,wherein the exogenous nucleic acid is incorporated into the DNA of the cell and expressed by the cell,wherein the method is performed without introducing nucleic acids into the cell that have specific binding to the protein-protein interaction domain.
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Abstract
Disclosed herein are methods and compositions for gene targeting utilizing fusion molecules comprising a recombinase domain and a sequence-specific DNA-binding domain.
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13 Claims
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1. A method of placing an exogenous nucleic acid into DNA of a cell comprising introducing into the cell:
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(i) an exogenous nucleic acid, (ii) a proteinaceous fusion molecule that comprises a recombinase domain, a protein-protein interaction domain, and a nuclear localization signal domain, and (iii) a single strand DNA probe complementary to a target site in DNA of the cell (cssDNA), with the probe and the recombinase domain complexed as a nucleoprotein filament at the time of the introduction into the cell, with the exogenous nucleic acid sequence being chosen for incorporation into the DNA of the cell and designed to be free of specific binding to the proteinaceous fusion molecule, wherein the exogenous nucleic acid is incorporated into the DNA of the cell and expressed by the cell, wherein the method is performed without introducing nucleic acids into the cell that have specific binding to the protein-protein interaction domain. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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Specification