Patent ID: 7164056
Filing Date: 2007-01-16
Classification: C12N

Abstract:
1. A method for gene targeting in a plant cell comprising: (a) providing the plant cell, wherein the plant cell comprises a target polynucleotide, wherein the target polynucleotide is endogenous to the plant cell; (b) introducing into the plant cell a targeting vector comprising a target modifying polynucleotide and a geminiviral origin of replication, wherein the target modifying polynucleotide has at least 75–150 bp of homology to the target polynucleotide, wherein the targeting vector is integrated into the plant genome, and wherein the targeting vector is flanked by site-specific recombination sites for excision; (c) providing a site-specific recombinase to excise the targeting vector, wherein the excised targeting vector forms a nucleic acid molecule capable of replication; (d) providing an appropriate viral replicase, wherein the replicase binds to the geminiviral origin of replication, to stimulate homologous recombination between the target modifying polynucleotide and the target polynucleotide to produce a modified target polynucleotide; and (e) recovering the plant cell comprising the modified target polynucleotide integrated into its genome.