Optimal soybean loci
First Claim
Patent Images
1. A recombinant soybean cell produced by the following steps:
- a. providing a soybean cell wherein said cell comprises a target nongenic soybean genomic nucleic acid molecule selected from the group consisting ofSEQ ID NO;
639, identified as soy_OGL_1423,SEQ ID NO;
137, identified as soy_OGL_1434,SEQ ID NO;
76, identified as soy_OGL_4625,SEQ ID NO;
440, identified as soy_OGL_6362,SEQ ID NO;
43, identified as soy_OGL_308,SEQ ID NO;
566, identified as soy_OGL_307,SEQ ID NO;
4236, identified as soy_OGL_310,SEQ ID NO;
47, identified as soy_OGL_684,SEQ ID NO;
2101, identified as soy_OGL_682, andSEQ ID NO;
48, identified as soy_OGL_685b. selecting a site specific nuclease that specifically binds and cleaves said target nongenic soybean genomic nucleic acid molecule;
c. introducing said site specific nuclease activity into said soybean cell;
d. introducing a DNA of interest into the soybean cell;
e. inserting the DNA of interest into said target nongenic soybean genomic nucleic acid molecule; and
,f. selecting transgenic soybean cells comprising the DNA of interest targeted to said nongenic nucleic acid molecule.
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Abstract
As disclosed herein, optimal native genomic loci of soybean plants have been identified that represent best sites for targeted insertion of exogenous sequences.
57 Citations
21 Claims
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1. A recombinant soybean cell produced by the following steps:
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a. providing a soybean cell wherein said cell comprises a target nongenic soybean genomic nucleic acid molecule selected from the group consisting of SEQ ID NO;
639, identified as soy_OGL_1423,SEQ ID NO;
137, identified as soy_OGL_1434,SEQ ID NO;
76, identified as soy_OGL_4625,SEQ ID NO;
440, identified as soy_OGL_6362,SEQ ID NO;
43, identified as soy_OGL_308,SEQ ID NO;
566, identified as soy_OGL_307,SEQ ID NO;
4236, identified as soy_OGL_310,SEQ ID NO;
47, identified as soy_OGL_684,SEQ ID NO;
2101, identified as soy_OGL_682, andSEQ ID NO;
48, identified as soy_OGL_685b. selecting a site specific nuclease that specifically binds and cleaves said target nongenic soybean genomic nucleic acid molecule; c. introducing said site specific nuclease activity into said soybean cell; d. introducing a DNA of interest into the soybean cell; e. inserting the DNA of interest into said target nongenic soybean genomic nucleic acid molecule; and
,f. selecting transgenic soybean cells comprising the DNA of interest targeted to said nongenic nucleic acid molecule. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. A soybean plant, soybean plant part, or soybean plant cell comprising a recombinant nucleic acid molecule, said recombinant nucleic acid molecule comprising
a nongenic nucleic acid molecule of at least 1 Kb, wherein a. the level of methylation of said nongenic nucleic acid molecule is 1% or less; -
b. the nongenic nucleic acid molecule shares less than 40% sequence identity with any other nucleic acid molecules contained in the soybean genome; c. the nongenic nucleic acid molecule is located within a 40 Kb region of a known or predicted expressive soybean coding nucleic acid molecule; and d. the nongenic nucleic acid molecule exhibits a recombination frequency within the soybean genome of greater than 0.01574 cM/Mb, wherein said nongenic nucleic acid molecule has at least 95% sequence identity with a nongenic nucleic acid molecule selected from the group consisting of SEQ ID NO;
639, identified as soy_OGL_1423,SEQ ID NO;
137, identified as soy_OGL_1434,SEQ ID NO;
76, identified as soy_OGL_4625,SEQ ID NO;
440, identified as soy_OGL_6362,SEQ ID NO;
43, identified as soy_OGL_308,SEQ ID NO;
566, identified as soy_OGL_307,SEQ ID NO;
4236, identified as soy_OGL_310,SEQ ID NO;
47, identified as soy_OGL_684,SEQ ID NO;
2101, identified as soy_OGL_682, andSEQ ID NO;
48, identified as soy_OGL_685; anda DNA of interest, wherein the DNA of interest comprises an open reading frame and said DNA of interest is inserted into said nongenic nucleic acid molecule to produce said recombinant nucleic acid molecule. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17)
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18. A soybean plant, soybean plant part, or soybean plant cell comprising a nucleic acid molecule encoding a site specific nuclease that specifically binds and cleaves a target nongenic soybean genomic nucleic acid molecule selected from the group consisting of
SEQ ID NO: - 639, identified as soy_OGL_1423,
SEQ ID NO;
137, identified as soy_OGL_1434,SEQ ID NO;
76, identified as soy_OGL_4625,SEQ ID NO;
440, identified as soy_OGL_6362,SEQ ID NO;
43, identified as soy_OGL_308,SEQ ID NO;
566, identified as soy_OGL_307,SEQ ID NO;
4236, identified as soy_OGL_310,SEQ ID NO;
47, identified as soy_OGL_684,SEQ ID NO;
2101, identified as soy_OGL_682, andSEQ ID NO;
48, identified as soy_OGL_685. - View Dependent Claims (19, 20, 21)
- 639, identified as soy_OGL_1423,
Specification