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US 12,729,383 B2Granted
Optimal soybean loci
AssigneeCorteva Agriscience LLC
InventorsLakshmi Sastry-Dent, Zehui Cao, Shreedharan Sriram, Steven R. Webb, Debra L. Camper, W. Michael Ainley
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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.
Claims19
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:
- 4106, identified as soy_ogl_275,
- SEQ ID NO:
- 4496, identified as soy_ogl_598,
- SEQ ID NO:
- 4875, identified as soy_ogl_5454,
- SEQ ID NO:
- 4888, identified as soy_ogl_6838,
- SEQ ID NO:
- 5063, identified as soy_ogl_4779,
- SEQ ID NO:
- 5687, identified as soy_ogl_796,
- SEQ ID NO:
- 6087, identified as soy_ogl_873, and
- SEQ ID NO:
- 6321, identified as soy_ogl_5475 and
- a DNA of interest, wherein the DNA of interest is inserted into said nongenic nucleic acid molecule to produce said recombinant nucleic acid molecule.
21 more elements
A method of making a transgenic plant cell comprising a DNA of interest, the method comprising:
- a. selecting a target nongenic soybean genomic nucleic acid molecule of at least 1 Kb, wherein
- i. the level of methylation of said nongenic nucleic acid molecule is 1% or less;
- ii. the nongenic nucleic acid molecule shares less than 40% sequence identity with any other nucleic acid molecule contained in the soybean genome;
- iii. the nongenic nucleic acid molecule is located within a 40 Kb region of a known or predicted expressive soybean coding nucleic acid molecule;
- and
- iv. 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 to a nucleic acid molecule selected from the group consisting of
- SEQ ID NO:
- 4106, identified as soy_ogl_275,
- SEQ ID NO:
- 4496, identified as soy_ogl_598,
- SEQ ID NO:
- 4875, identified as soy_ogl_5454,
- SEQ ID NO:
- 4888, identified as soy_ogl_6838,
- SEQ ID NO:
- 5063, identified as soy_ogl_4779,
- SEQ ID NO:
- 5687, identified as soy_ogl_796,
- SEQ ID NO:
- 6087, identified as soy_ogl_873, and
- SEQ ID NO:
- 6321, identified as soy_ogl_5475;
- b. selecting a site specific nuclease that specifically binds and cleaves said target nongenic soybean genomic nucleic acid molecule;
- c. introducing said site specific nuclease into a soybean plant cell;
- d. introducing the DNA of interest into the plant cell;
- e. inserting the DNA of interest into said target nongenic soybean genomic nucleic acid molecule;
- and,
- f. selecting transgenic plant cells comprising the DNA of interest inserted into said nongenic nucleic acid molecule.
26 more elements
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 having at least 95% sequence identity to a nucleic acid molecule selected from the group consisting of
- SEQ ID NO:
- 4106, identified as soy_ogl_275,
- SEQ ID NO:
- 4496, identified as soy_ogl_598,
- SEQ ID NO:
- 4875, identified as soy_ogl_5454,
- SEQ ID NO:
- 4888, identified as soy_ogl_6838,
- SEQ ID NO:
- 5063, identified as soy_ogl_4779,
- SEQ ID NO:
- 5687, identified as soy_ogl_796,
- SEQ ID NO:
- 6087, identified as soy_ogl_873, and
- SEQ ID NO:
- 6321, identified as soy_ogl_5475;
- b. 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 inserted into said nongenic nucleic acid molecule.
20 more elements
Dependencies are inferred from the claim text.