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Last Updated: April 1, 2020

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Claims for Patent: 7,901,905

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Summary for Patent: 7,901,905
Title:Gene expression system based on codon translation efficiency
Abstract: The present invention discloses a method for modulating the production of a protein from a polynucleotide in a CHO cell by replacing at least one codon of the polynucleotide with a synonymous codon that has a higher or lower translation efficiency in the CHO cell than the codon it replaces, or by introducing into the CHO cell a polynucleotide that codes for an iso-tRNA which limits the rate of production of the polypeptide and which corresponds to a codon of the first polynucleotide. The present invention also discloses the use of a protein-encoding polynucleotide whose codon composition has been modified for enhanced production of the protein in CHO cells.
Inventor(s): Frazer; Ian Hector (Queensland, AU)
Assignee: The University of Queensland (St. Lucia, Queensland, AU)
Application Number:11/077,939
Patent Claims:1. A method of constructing a synthetic polynucleotide from which a polypeptide is producible at a higher level in a Chinese Hamster Ovary (CHO) cell than from a parent polynucleotide encoding the same polypeptide, the method comprising: selecting a first codon of the parent polynucleotide for replacement with a synonymous codon, wherein the synonymous codon is selected on the basis that it exhibits a higher translational efficiency in the CHO cell than the first codon in a comparison of translational efficiencies of codons in test CHO cells; and replacing the first codon with the synonymous codon to construct the synthetic polynucleotide, wherein the first and synonymous codons are selected from the following Table: TABLE-US-00013 Synonymous First Codon Codon Ala.sup.GCC Ala.sup.GCA Ala.sup.GCC Ala.sup.GCG Ala.sup.GCC Ala.sup.GCT Ala.sup.GCT Ala.sup.GCA Ala.sup.GCG Ala.sup.GCA Arg.sup.CGC Arg.sup.AGA Arg.sup.CGC Arg.sup.CGA Arg.sup.CGC Arg.sup.CGG Arg.sup.CGC Arg.sup.CGT Arg.sup.CGC Arg.sup.AGG Asp.sup.GAC Asp.sup.GAT Glu.sup.GAG Glu.sup.GAA Gly.sup.GGT Gly.sup.GGA Gly.sup.GGC Gly.sup.GGA Gly.sup.GGC Gly.sup.GGG Gly.sup.GGG Gly.sup.GGA Leu.sup.CTT Leu.sup.TTG Leu.sup.CTG Leu.sup.CTC Leu.sup.CTG Leu.sup.TTG Leu.sup.CTA Leu.sup.TTG Leu.sup.CTT Leu.sup.CTA Leu.sup.TTA Leu.sup.CTA Phe.sup.TTC Phe.sup.TTT Thr.sup.ACC Thr.sup.ACT Thr.sup.ACC Thr.sup.ACG Thr.sup.ACC Thr.sup.ACA.

2. A method according to claim 1, wherein the polypeptide is selected from the group consisting of etanercept, HPV16E7 and human growth hormone.

3. A method of producing a polypeptide in a Chinese Hamster Ovary (CHO) cell from a synthetic polynucleotide at a higher level than from a parent polynucleotide encoding the same polypeptide, the method comprising: selecting a first codon of the parent polynucleotide for replacement with a synonymous codon, wherein the synonymous codon is selected on the basis that it exhibits a higher translational efficiency in the CHO cell than the first codon in a comparison of translational efficiencies of codons in test CHO cells, wherein both the first and synonymous codons are selected from the following Table: TABLE-US-00014 Synonymous First Codon Codon Ala.sup.GCC Ala.sup.GCA Ala.sup.GCC Ala.sup.GCG Ala.sup.GCC Ala.sup.GCT Ala.sup.GCT Ala.sup.GCA Ala.sup.GCG Ala.sup.GCA Arg.sup.CGC Arg.sup.AGA Arg.sup.CGC Arg.sup.CGA Arg.sup.CGC Arg.sup.CGG Arg.sup.CGC Arg.sup.CGT Arg.sup.CGC Arg.sup.AGG Asp.sup.GAC Asp.sup.GAT Glu.sup.GAG Glu.sup.GAA Gly.sup.GGT Gly.sup.GGA Gly.sup.GGC Gly.sup.GGA Gly.sup.GGC Gly.sup.GGG Gly.sup.GGG Gly.sup.GGA Leu.sup.CTT Leu.sup.TTG Leu.sup.CTG Leu.sup.CTC Leu.sup.CTG Leu.sup.TTG Leu.sup.CTA Leu.sup.TTG Leu.sup.CTT Leu.sup.CTA Leu.sup.TTA Leu.sup.CTA Phe.sup.TTC Phe.sup.TTT Thr.sup.ACC Thr.sup.ACT Thr.sup.ACC Thr.sup.ACG Thr.sup.ACC Thr.sup.ACA

replacing the first codon with the synonymous codon to construct the synthetic polynucleotide; introducing the synthetic polynucleotide into the CHO cell; and expressing the synthetic polynucleotide in the CHO cell, whereby the polypeptide is produced from the synthetic polynucleotide in the CHO cell at a higher level than from the parent polynucleotide.

4. A method according to claim 3, further comprising isolating or purifying the polypeptide from the CHO cell.

5. A method of producing a virus particle in a Chinese Hamster Ovary (CHO) cell, wherein the virus particle comprises a polypeptide necessary for assembly of the virus particle, and wherein the polypeptide is produced in the CHO cell from a parent polynucleotide, but not at a level sufficient to permit productive virus assembly therein, the method comprising: selecting a first codon of the parent polynucleotide for replacement with a synonymous codon, wherein the synonymous codon is selected on the basis that it exhibits a higher translational efficiency in the CHO cell than the first codon in a comparison of translational efficiencies of codons in test CHO cells, wherein both the first and synonymous codons are selected from the following Table: TABLE-US-00015 Synonymous First Codon Codon Ala.sup.GCC Ala.sup.GCA Ala.sup.GCC Ala.sup.GCG Ala.sup.GCC Ala.sup.GCT Ala.sup.GCT Ala.sup.GCA Ala.sup.GCG Ala.sup.GCA Arg.sup.CGC Arg.sup.AGA Arg.sup.CGC Arg.sup.CGA Arg.sup.CGC Arg.sup.CGG Arg.sup.CGC Arg.sup.CGT Arg.sup.CGC Arg.sup.AGG Asp.sup.GAC Asp.sup.GAT Glu.sup.GAG Glu.sup.GAA Gly.sup.GGT Gly.sup.GGA Gly.sup.GGC Gly.sup.GGA Gly.sup.GGC Gly.sup.GGG Gly.sup.GGG Gly.sup.GGA Leu.sup.CTT Leu.sup.TTG Leu.sup.CTG Leu.sup.CTC Leu.sup.CTG Leu.sup.TTG Leu.sup.CTA Leu.sup.TTG Leu.sup.CTT Leu.sup.CTA Leu.sup.TTA Leu.sup.CTA Phe.sup.TTC Phe.sup.TTT Thr.sup.ACC Thr.sup.ACT Thr.sup.ACC Thr.sup.ACG Thr.sup.ACC Thr.sup.ACA

replacing the first codon with the synonymous codon to construct the synthetic polynucleotide; and introducing into the CHO cell the synthetic polynucleotide operably linked to a regulatory polynucleotide, whereby the synthetic polynucleotide is expressed to produce the polypeptide at a level sufficient to permit the production of the virus particle in the CHO cell.

6. A method according to claim 5, further comprising isolating or purifying the virus particle from the CHO cell.

Details for Patent 7,901,905

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Immunex ENBREL etanercept VIAL; SUBCUTANEOUS 103795 001 1998-11-02   Start Trial The University of Queensland (St. Lucia, Queensland, AU) 2022-09-13 RX Orphan search
Immunex ENBREL etanercept SYRINGE 103795 002 1998-11-02   Start Trial The University of Queensland (St. Lucia, Queensland, AU) 2022-09-13 RX Orphan search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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