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Method to enhance homologous recombination

  • US 20030228608A1
  • Filed: 04/15/2003
  • Published: 12/11/2003
  • Est. Priority Date: 04/16/2002
  • Status: Abandoned Application
First Claim
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1. A method for targeting and altering, by homologous recombination, a preselected target nucleic acid sequence in an extrachromosomal sequence, comprising:

  • a) providing a mixture comprising recombinase and an at least partially single stranded nucleic acid substrate for recombination comprising two nucleic acid molecules, wherein the first and the second nucleic acid molecules each comprise targeting polynucleotides which substantially correspond to or are substantially complementary to the preselected target nucleic acid sequence, wherein the two nucleic acid molecules are capable of forming a partially double stranded molecule with each other, and wherein at least the 5′

    end or the 3′

    end of the first nucleic acid molecule comprises a nucleotide sequence, the substantial complement of which is not present at the 3′

    end or 5′

    end of the second nucleic acid molecule, which nucleotide sequence is capable of binding recombinase;

    b) contacting the mixture with the extrachromosomal sequence to form a recombination intermediate; and

    c) introducing the recombination intermediate into a cell to yield an altered cell comprising a genetically altered extrachromosomal sequence comprising a targeted sequence alteration.

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