Precision gene targeting to a particular locus in maize
First Claim
1. A method for integrating one or more exogenous nucleic acid sequences into the genome of a maize cell having an E32 locus, said method comprising the steps of:
- making a double-stranded cleavage in the E32 locus using a site specific zinc finger nuclease, wherein the site specific zinc finger nuclease comprises three or more zinc finger motifs selected from the group consisting of SEQ ID NO;
43, SEQ ID NO;
44, SEQ ID NO;
45, SEQ ID NO;
46, SEQ ID NO;
47, SEQ ID NO;
48, SEQ ID NO;
49, SEQ ID NO;
50, SEQ ID NO;
51, and SEQ ID NO;
52; and
ligating a polynucleotide comprising one or more exogenous nucleic acid sequences into the cleavage using a ligase or a recombinase, wherein the polynucleotide is integrated into the cleavage.
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Abstract
The present invention claims methods for the stable integration of exogenous DNA into a specific locus, E32, in the maize genome through the use of zinc finger nucleases. Maize plants and plant parts that were transformed by the methods of the invention are claimed. The invention is useful for creating desirable traits such as herbicide resistance, herbicide tolerance, insect resistance, insect tolerance, disease resistance, disease tolerance, stress tolerance, and stress resistance in maize The E32 locus represents a superior site for inserting foreign genes because native agronomic phenotypes are not disturbed.
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Citations
13 Claims
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1. A method for integrating one or more exogenous nucleic acid sequences into the genome of a maize cell having an E32 locus, said method comprising the steps of:
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making a double-stranded cleavage in the E32 locus using a site specific zinc finger nuclease, wherein the site specific zinc finger nuclease comprises three or more zinc finger motifs selected from the group consisting of SEQ ID NO;
43, SEQ ID NO;
44, SEQ ID NO;
45, SEQ ID NO;
46, SEQ ID NO;
47, SEQ ID NO;
48, SEQ ID NO;
49, SEQ ID NO;
50, SEQ ID NO;
51, and SEQ ID NO;
52; andligating a polynucleotide comprising one or more exogenous nucleic acid sequences into the cleavage using a ligase or a recombinase, wherein the polynucleotide is integrated into the cleavage. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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Specification