Optimal soybean loci
First Claim
Patent Images
1. A soybean plant, soybean plant part, or soybean plant cell comprising a recombinant nucleic acid molecule, said recombinant nucleic acid molecule comprisinga nongenic soybean genomic sequence of at least 1 Kb determined to comprise the following characteristics:
- a. the level of methylation of said nongenic sequence is 1% or less;
b. said nongenic sequence shares less than 40% sequence identity with any other sequence contained in the Glycine max genome;
c. said nongenic sequence is located within a 40 Kb region of a known or predicted expressive soybean coding sequence; and
d. said nongenic sequence exhibits a recombination frequency within the soybean genome of greater than 0.01574 cM/Mb;
wherein said nongenic sequence comprises a 1 Kb sequence having at least 95% sequence identity with a sequence selected from the group consisting of SEQ ID NO;
2639, SEQ ID NO;
3682, SEQ ID NO;
4050, SEQ ID NO;
4593, SEQ ID NO;
4622, SEQ ID NO;
4879, SEQ ID NO;
4932, SEQ ID NO;
5102, SEQ ID NO;
5122, SEQ ID NO;
5520, SEQ ID NO;
5698, SEQ ID NO;
6087, SEQ ID NO;
6515, SEQ ID NO;
6571, SEQ ID NO;
6586, SEQ ID NO;
6775 and SEQ ID NO;
6935; and
a DNA of interest, wherein the DNA of interest is inserted into said nongenic sequence.
3 Assignments
0 Petitions
Accused Products
Abstract
As disclosed herein, optimal native genomic loci have been identified in dicot plants, such as soybean plants, that represent best sites for targeted insertion of exogenous sequences.
60 Citations
9 Claims
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1. A soybean plant, soybean plant part, or soybean plant cell comprising a recombinant nucleic acid molecule, said recombinant nucleic acid molecule comprising
a nongenic soybean genomic sequence of at least 1 Kb determined to comprise the following characteristics: -
a. the level of methylation of said nongenic sequence is 1% or less; b. said nongenic sequence shares less than 40% sequence identity with any other sequence contained in the Glycine max genome; c. said nongenic sequence is located within a 40 Kb region of a known or predicted expressive soybean coding sequence; and d. said nongenic sequence exhibits a recombination frequency within the soybean genome of greater than 0.01574 cM/Mb;
wherein said nongenic sequence comprises a 1 Kb sequence having at least 95% sequence identity with a sequence selected from the group consisting of SEQ ID NO;
2639, SEQ ID NO;
3682, SEQ ID NO;
4050, SEQ ID NO;
4593, SEQ ID NO;
4622, SEQ ID NO;
4879, SEQ ID NO;
4932, SEQ ID NO;
5102, SEQ ID NO;
5122, SEQ ID NO;
5520, SEQ ID NO;
5698, SEQ ID NO;
6087, SEQ ID NO;
6515, SEQ ID NO;
6571, SEQ ID NO;
6586, SEQ ID NO;
6775 and SEQ ID NO;
6935; anda DNA of interest, wherein the DNA of interest is inserted into said nongenic sequence. - View Dependent Claims (2, 3, 4, 5)
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6. A method of making a transgenic plant cell comprising a DNA of interest targeted to one nongenic soybean genomic sequence, the method comprising:
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a. selecting an optimal nongenic soybean genomic sequence of at least 1 Kb, wherein said nongenic soybean genomic sequence comprises the following characteristics; ii). the level of methylation of said nongenic sequence is 1% or less; ii). said nongenic sequence shares less than 40% sequence identity with any other sequence contained in the Glycine max genome; iii). said nongenic sequence is located within a 40 Kb region of a known or predicted expressive soybean coding sequence; and iv). said nongenic sequence exhibits a recombination frequency within the soybean genome of greater than 0.01574 cM/Mb;
further wherein said nongenic sequence comprises a 1 Kb sequence having at least 95% sequence identity with a sequence selected from the group consisting of SEQ ID NO;
2639, SEQ ID NO;
3682, SEQ ID NO;
4050, SEQ ID NO;
4593, SEQ ID NO;
4622, SEQ ID NO;
4879, SEQ ID NO;
4932, SEQ ID NO;
5102, SEQ ID NO;
5122, SEQ ID NO;
5520, SEQ ID NO;
5698, SEQ ID NO;
6087, SEQ ID NO;
6515, SEQ ID NO;
6571, SEQ ID NO;
6586, SEQ ID NO;
6775 and SEQ ID NO;
6935;b. introducing a site specific nuclease into a plant cell, wherein the site specific nuclease cleaves said nongenic soybean 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 (7, 8, 9)
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Specification