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US 11,441,151 B2Granted
Increased protein expression in plants
AssigneeDow AgroSciences LLC
InventorsAaron Woosley, Sarah Worden
Life of this patent
Estimating the term…
Abstract
This disclosure concerns synthetic polynucleotides encoding a polypeptide of interest that are particularly well-suited for expression in target plants.
Claims24
A polynucleotide comprising:
- a plant promoter that is functional in maize;
- and
- a synthetic nucleotide sequence encoding a polypeptide encoded by a native nucleotide sequence in the genome of an organism other than a maize plant,
- wherein the synthetic nucleotide sequence has been codon-optimized to remove codons selected from the group consisting of TTA, CTA, GTA, CGT, AGT, and CGA, from the native nucleotide sequence,
- wherein the synthetic nucleotide sequence comprises at least one polyadenylation sequence selected from the group consisting of AATAAA, AATAAT, AACCAA, ATATAA, AATCAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA, present in the same number and in the same location as in the native nucleotide sequence,
- wherein the synthetic nucleotide sequence does not comprise any polyadenylation sequence selected from the group consisting of AATAAT, AACCAA, ATATAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA in a different number or location than in the native nucleotide sequence, and
- wherein the synthetic nucleotide sequence does not comprise a polyadenylation sequence selected from the group consisting of ATATAT, TTGTTT, TTTTGT, TGTTTT, TATATA, TATTTT, TTTTTT, ATTTTT, TTATTT, TTTATT, TAATAA, ATTTAT, TATATT, TTTTAT, ATATTT, TATTAT, TGTTTG, TTATAT, TGTAAT, AAATAA, AATTTT, TTTTTA, TAATTT, TTAATT, AAATTT, TTTGTT, ATTATT, ATTTTA, TTTAAT, and TTTTAA.
4 more elements
A method for producing a synthetic polynucleotide, the method comprising:
- removing codons selected from the group consisting of TTA, CTA, GTA, CGT, AGT, and CGA, from a nucleotide sequence encoding a polypeptide that is native in an organism other than a maize plant, to produce a codon-optimized nucleotide sequence;
- engineering the codon-optimized nucleotide sequence such that:
- the codon-optimized nucleotide sequence comprises at least one polyadenylation sequence selected from the group consisting of AATAAA, AATAAT, AACCAA, ATATAA, AATCAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA, present in the same number and in the same location as in the nucleotide sequence encoding the polypeptide that is native in the organism other than a maize plant,
- the codon-optimized nucleotide sequence does not comprise any polyadenylation sequence selected from the group consisting of AATAAT, AACCAA, ATATAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA in a different number or location than in the nucleotide sequence encoding the polypeptide that is native in the organism other than a maize plant, and
- the codon-optimized nucleotide sequence does not comprise a polyadenylation sequence selected from the group consisting of ATATAT, TTGTTT, TTTTGT, TGTTTT, TATATA, TATTTT, TTTTTT, ATTTTT, TTATTT, TTTATT, TAATAA, ATTTAT, TATATT, TTTTAT, ATATTT, TATTAT, TGTTTG, TTATAT, TGTAAT, AAATAA, AATTTT, TTTTTA, TAATTT, TTAATT, AAATTT, TTTGTT, ATTATT, ATTTTA, TTTAAT, and TTTTAA;
- and
- constructing the synthetic polynucleotide.
4 more elements
A polynucleotide comprising:
- a plant promoter that is functional in soybean;
- and
- a synthetic nucleotide sequence encoding a polypeptide encoded by a native nucleotide sequence in the genome of an organism other than a soybean plant,
- wherein the synthetic nucleotide sequence has been codon-optimized to remove codons selected from the group consisting of TTA, CTA, TCG, CCG, ACG, GCG, CGA, and CGG, from the native nucleotide sequence,
- wherein the synthetic nucleotide sequence comprises at least one polyadenylation sequence selected from the group consisting of AATAAA, AATAAT, AACCAA, ATATAA, AATCAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA, present in the same number and in the same location as in the native nucleotide sequence,
- wherein the synthetic nucleotide sequence does not comprise any polyadenylation sequence selected from the group consisting of AATAAT, AACCAA, ATATAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA in a different number or location than in the native nucleotide sequence, and
- wherein the synthetic nucleotide sequence does not comprise a polyadenylation sequence selected from the group consisting of ATATAT, TTGTTT, TTTTGT, TGTTTT, TATATA, TATTTT, TTTTTT, ATTTTT, TTATTT, TTTATT, TAATAA, ATTTAT, TATATT, TTTTAT, ATATTT, TATTAT, TGTTTG, TTATAT, TGTAAT, AAATAA, AATTTT, TTTTTA, TAATTT, TTAATT, AAATTT, TTTGTT, ATTATT, ATTTTA, TTTAAT, and TTTTAA.
4 more elements
A method for producing a synthetic polynucleotide, the method comprising:
- removing codons selected from the group consisting of TTA, CTA, TCG, CCG, ACG, GCG, CGA, and CGG, from a nucleotide sequence encoding a polypeptide that is native in an organism other than a soybean plant, to produce a codon-optimized nucleotide sequence;
- engineering the codon-optimized nucleotide sequence such that:
- the codon-optimized nucleotide sequence comprises at least one polyadenylation sequence selected from the group consisting of AATAAA, AATAAT, AACCAA, ATATAA, AATCAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA, present in the same number and in the same location as in the nucleotide sequence encoding the polypeptide that is native in the organism other than a maize plant,
- the codon-optimized nucleotide sequence does not comprise any polyadenylation sequence selected from the group consisting of AATAAT, AACCAA, ATATAA, ATACTA, ATAAAA, ATGAAA, AAGCAT, ATTAAT, ATACAT, AAAATA, ATTAAA, AATTAA, AATACA, and CATAAA in a different number or location than in the nucleotide sequence encoding the polypeptide that is native in the organism other than a maize plant, and
- the codon-optimized nucleotide sequence does not comprise a polyadenylation sequence selected from the group consisting of ATATAT, TTGTTT, TTTTGT, TGTTTT, TATATA, TATTTT, TTTTTT, ATTTTT, TTATTT, TTTATT, TAATAA, ATTTAT, TATATT, TTTTAT, ATATTT, TATTAT, TGTTTG, TTATAT, TGTAAT, AAATAA, AATTTT, TTTTTA, TAATTT, TTAATT, AAATTT, TTTGTT, ATTATT, ATTTTA, TTTAAT, and TTTTAA;
- and
- constructing the synthetic polynucleotide.
4 more elements
Dependencies are inferred from the claim text.