Optimal Maize Loci
First Claim
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1. A recombinant sequence, said recombinant sequence comprising:
- a nongenic maize genomic sequence of at least 1 Kb, said nongenic sequence being hypomethylated, targetable, located proximal to a genic region within a maize genome, and exemplifying evidence of recombination; and
an DNA of interest, wherein the DNA of interest is inserted into said nongenic sequence.
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Abstract
As disclosed herein, optimal native genomic loci have been identified in monocot plants, such as maize plants, that represent best sites for targeted insertion of exogenous sequences.
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Citations
23 Claims
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1. A recombinant sequence, said recombinant sequence comprising:
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a nongenic maize genomic sequence of at least 1 Kb, said nongenic sequence being hypomethylated, targetable, located proximal to a genic region within a maize genome, and exemplifying evidence of recombination; and an DNA of interest, wherein the DNA of interest is inserted into said nongenic sequence. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A method of making a transgenic plant cell comprising a DNA of interest targeted to one nongenic maize genomic sequence, the method comprising:
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a. selecting an optimal nongenic maize genomic locus; b. introducing a site specific nuclease into a plant cell, wherein the site specific nuclease cleaves said nongenic maize genomic sequence; c. introducing the DNA of interest into the plant cell; d. targeting the DNA of interest into said nongenic locus, wherein the cleavage of said nongenic sequence facilitates integration of the polynucleotide sequence into said nongenic locus; and e. selecting transgenic plant cells comprising the DNA of interest targeted to said nongenic locus. - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 22)
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23. A recombinant targetable nongenic maize genomic sequence comprising:
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a nongenic sequence of at least 1 Kb selected from the group consisting of cluster 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 or a sequence sharing 99% sequence identity with a sequence selected from the group consisting of cluster 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, and their respective complements thereof, wherein said clusters are generated from a PCA statistical algorithm comprising PCA values defined in Table 6; and a DNA of interest inserted into said nongenic sequence.
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Specification