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Methods for the replacement, translocation and stacking of DNA in eukaryotic genomes

  • US 20020123145A1
  • Filed: 07/23/2001
  • Published: 09/05/2002
  • Est. Priority Date: 07/21/2000
  • Status: Active Grant
First Claim
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1. A method of obtaining site-specific gene replacement in a eukaryotic cell, comprising:

  • a) providing a eukaryotic cell that comprises a receptor construct, wherein the receptor construct comprises a receptor polynucleotide flanked by two or more of a irreversible recombination site (IRS);

    b) introducing into the cell a donor construct that comprises a donor polynucleotide flanked by two or more of a irreversible complementary recombination site (CIRS); and

    c) contacting the receptor construct and the donor construct with an irreversible recombinase polypeptide;

    d) wherein the irreversible recombinase catalyzes recombination between the first and second types of recombination sites and replacement of the receptor polynucleotide with the donor polynucleotide, thereby forming a replacement construct.

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