EXCISION OF TRANSGENES IN GENETICALLY MODIFIED ORGANISMS

摘要

A method for deleting a region of DNA in a plant. In some embodiments, the method comprises transforming a plant with a nucleic acid molecule, wherein the nucleic acid molecule encodes one or more zinc finger nuclease(s) (ZFNs) operably linked to one or more tissue-specific promoter(s), e.g., a pollen-specific promoter. Methods include excising native genes in a plant. Accordingly, in some embodiments, ZFNs are engineered that recognize sequences that flank native plant genes. In further embodiments, ZFNs are expressed under the control of developmental stage-specific promoters, such that, for example, nucleic acid sequences are specifically excised in plants during relatively late stages of development. Nucleic acid molecules useful for carrying out disclosed methods and plants produced by the methods are included.

主权项

1. A method for segregating a first polynucleotide fragment from a second polynucleotide fragment within a genome of a plant cell, wherein the first polynucleotide fragment and the second polynucleotide fragment are located in close proximity on a single chromosome, the method comprising the steps of:
(a) providing the plant cell comprising the first polynucleotide fragment and the second polynucleotide fragment located in close proximity on the single chromosome, the plant cell further comprising at least a second chromosome, wherein the first and second chromosomes are homologous or homeologous chromosomes of each other; (b) introducing a site specific nuclease into the genome of the plant cell; (c) producing a double strand break in the first chromosome, wherein the double strand break in the first chromosome is introduced between the first polynucleotide fragment and the second polynucleotide fragment; (d) producing a double strand break in the second chromosome, wherein the double strand break in the second chromosome is introduced between the first polynucleotide fragment and the second polynucleotide fragment; and (e) obtaining a progeny plant comprising a modified genome, wherein the first polynucleotide fragment and the second polynucleotide fragment segregate from one another.