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US 12,735,716 B2Granted
Optimal maize loci
AssigneeCorteva Agriscience LLC
InventorsLakshmi Sastry-Dent, Zehui Cao, Shreedharan Sriram, Steven R. Webb, Debra L. Camper
Life of this patent
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Abstract
As disclosed herein, optimal native genomic loci from maize plants have been identified that represent best sites for targeted insertion of exogenous sequences.
Claims14
A maize plant, maize plant part, or maize 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 maize genome;
- c. the nongenic nucleic acid molecule is located within a 40 Kb region of a known or predicted expressive maize coding nucleic acid molecule;
- and
- d. the nongenic nucleic acid molecule exhibits a recombination frequency within the maize genome of greater than 0.00041 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:199,
- SEQ ID NO:365,
- SEQ ID NO:543,
- SEQ ID NO:687,
- SEQ ID NO:1089,
- SEQ ID NO:1233,
- SEQ ID NO:1369, and
- SEQ ID NO:1795;
- and
- a DNA of interest, wherein the DNA of interest comprises a non-native exogenous sequence, and said DNA of interest is inserted into said nongenic nucleic acid molecule to produce said recombinant nucleic acid molecule.
13 more elements
A maize plant, maize plant part, or maize 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 maize genome;
- c. the nongenic nucleic acid molecule is located within a 40 Kb region of a known or predicted expressive maize coding nucleic acid molecule;
- and
- d. the nongenic nucleic acid molecule exhibits a recombination frequency within the maize genome of greater than 0.00041 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:199,
- SEQ ID NO:365,
- SEQ ID NO:543,
- SEQ ID NO:687,
- SEQ ID NO:1089,
- SEQ ID NO:1233,
- SEQ ID NO:1369, and
- SEQ ID NO:1795;
- and
- a DNA of interest, wherein said DNA of interest comprises two or more gene expression cassettes and said DNA of interest is inserted into said nongenic nucleic acid molecule to produce said recombinant nucleic acid molecule.
13 more elements
A method of making a transgenic plant cell comprising a DNA of interest, the method comprising:
- a. selecting a target nongenic maize 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 maize genome;
- iii, the nongenic nucleic acid molecule is located within a 40 Kb region of a known or predicted expressive maize coding nucleic acid molecule;
- and
- iv. the nongenic nucleic acid molecule exhibits a recombination frequency within the maize genome of greater than 0.00041 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:199,
- SEQ ID NO:365,
- SEQ ID NO:543,
- SEQ ID NO:687,
- SEQ ID NO:1089,
- SEQ ID NO:1233,
- SEQ ID NO:1369, and
- SEQ ID NO:1795;
- and
- b. selecting a site specific nuclease that specifically binds and cleaves said target nongenic maize genomic nucleic acid molecule;
- c. introducing said site specific nuclease into a maize plant cell;
- d. introducing the DNA of interest into the plant cell;
- e. inserting the DNA of interest into said target nongenic maize genomic nucleic acid molecule;
- and,
- f. selecting transgenic plant cells comprising the DNA of interest targeted to said nongenic nucleic acid molecule.
18 more elements
Dependencies are inferred from the claim text.