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Last Updated: March 29, 2024

Claims for Patent: 8,114,628


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Summary for Patent: 8,114,628
Title:Expanding the eukaryotic genetic code
Abstract: This invention provides compositions and methods for producing translational components that expand the number of genetically encoded amino acids in eukaryotic cells. The components include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, orthogonal pairs of tRNAs/synthetases and unnatural amino acids. Proteins and methods of producing proteins with unnatural amino acids in eukaryotic cells are also provided.
Inventor(s): Chin; Jason W. (Cambridge, GB), Cropp; T. Ashton (Bethesda, MD), Anderson; J. Christopher (San Francisco, CA), Schultz; Peter G. (La Jolla, CA)
Assignee: The Scripps Research Institute (La Jolla, CA)
Application Number:11/978,121
Patent Claims:1. A eukaryotic cell comprising an orthogonal aminoacyl-tRNA synthetase (O-RS) as set forth in any one of SEQ ID NO: 36-47, or 86, wherein the O-RS preferentially aminoacylates an orthogonal tRNA (O-tRNA) with at least one unnatural amino acid in the eukaryotic cell wherein said unnatural amino acid is selected from a p-acetyl-L-phenylalanine, a p-iodo-L-phenylalanine, an O-methyl-L-tyrosine, or a p-benzoyl-L-phenylalanine.

2. The cell of claim 1, wherein the at least one unnatural amino acid comprises two or more unnatural amino acids.

3. The cell of claim 1, wherein the O-RS, or a portion thereof, is encoded by a polynucleotide sequence as set forth in any one of SEQ ID NO: 3-19.

4. The cell of claim 1, wherein the O-RS aminoacylates the O-tRNA with the at least one unnatural amino acid at least 10-fold more efficiently than the O-RS aminoacylates the O-tRNA with a natural amino acid.

5. The cell of claim 1, wherein the O-RS is derived from a non-eukaryotic organism.

6. The cell of claim 5, wherein the non-eukaryotic organism is Escherichia coli, or Bacillus stearothermophilus.

7. The cell of claim 1, wherein the eukaryotic cell is a yeast cell, a mammalian cell, a plant cell, an algae cell, a fungal cell, or an insect cell.

8. The cell of claim 7, wherein the eukaryotic cell is a Saccharomyces cerevisiae cell or a Pichia pastoris cell.

9. The cell of claim 8, wherein the O-tRNA is derived from a non-eukaryotic organism.

10. The cell of claim 9, wherein the non-eukaryotic organism is Escherichia coli, or Bacillus stearothermophilus.

11. The cell of claim 8, further comprising a nucleic acid that comprises a polynucleotide that encodes a polypeptide of interest, wherein the polynucleotide comprises a selector codon that is recognized by the O-tRNA.

12. The cell of claim 11, wherein the polypeptide of interest comprises a protein or a portion of a protein selected from the group consisting of: a cytokine, a growth factor, a growth factor receptor, an interferon, an interleukin, an inflammatory molecule, an oncogene product, a peptide hormone, a signal transduction molecule, a steroid hormone receptor, erythropoietin (EPO), insulin, human growth hormone, an Alpha-1 antitrypsin, an Angiostatin, an Antihemolytic factor, an antibody, an Apolipoprotein, an Apoprotein, an Atrial natriuretic factor, an Atrial natriuretic polypeptide, an Atrial peptide, a C--X--C chemokine, T39765, NAP-2, ENA-78, a Gro-a, a Gro-b, a Gro-c, an IP-10, a GCP-2, an NAP-4, an SDF-1, a PF4, a MIG, a Calcitonin, a c-kit ligand, a cytokine, a CC chemokine, a Monocyte chemoattractant protein-1, a Monocyte chemoattractant protein-2, a Monocyte chemoattractant protein-3, a Monocyte inflammatory protein-1 alpha, a Monocyte inflammatory protein-1 beta, RANTES, 1309, R83915, R91733, HCC1, T58847, D31065, T64262, a CD40, a CD40 ligand, a C-kit Ligand, a Collagen, a Colony stimulating factor (CSF), a Complement factor 5a, a Complement inhibitor, a Complement receptor 1, a cytokine, DHFR, an epithelial Neutrophil Activating Peptide-78, a GRO.alpha./MGSA, a GRO.beta., a GRO.gamma. a MIP-1.alpha., a MIP-1.delta., a MCP-1, an Epidermal Growth Factor (EGF), an epithelial Neutrophil Activating Peptide, an Erythropoietin (EPO), an Exfoliating toxin, a Factor IX, a Factor VII, a Factor VIII, a Factor X, a Fibroblast Growth Factor (FGF), a Fibrinogen, a Fibronectin, a G-CSF, a GM-CSF, a Glucocerebrosidase, a Gonadotropin, a growth factor, a growth factor receptor, a Hedgehog protein, a Hemoglobin, a Hepatocyte Growth Factor (HGF), a Hirudin, a Human serum albumin, an ICAM-1, an ICAM-1 receptor, an LFA-1, an LFA-1 receptor, an Insulin, an Insulin-like Growth Factor (IGF), an IGF-I, an IGF-II, an interferon, an IFN-.alpha., an IFN-.beta., an IFN-.gamma., an interleukin, an IL-1, an IL-2, an IL-3, an IL-4, an IL-5, an IL-6, an IL-7, an IL-8, an IL-9, an IL-10, an IL-11, an IL-12, a Keratinocyte Growth Factor (KGF), a Lactoferrin, a leukemia inhibitory factor, a Luciferase, a Neurturin, a Neutrophil inhibitory factor (NIF), an oncostatin M, an Osteogenic protein, an oncogene product, a Parathyroid hormone, a PD-ECSF, a PDGF, a peptide hormone, a Human Growth Hormone, a Pleiotropin, a Protein A, a Protein G, a Pyrogenic exotoxins A, B, or C, a Relaxin, a Renin, an SCF, a Soluble complement receptor I, a Soluble I-CAM 1, a Soluble interleukin receptor, a Soluble TNF receptor, a Somatomedin, a Somatostatin, a Somatotropin, a Streptokinase, a Superantigen, a Staphylococcal enterotoxins, an SEA, an SEB, an SEC 1, an SEC2, an SEC3, an SED, an SEE, a steroid hormone receptor, a Superoxide dismutase (SOD), a Toxic shock syndrome toxin, a Thymosin alpha 1, a Tissue plasminogen activator, a tumor growth factor (TGF), a TGF-.alpha., a TGF-.beta., a Tumor Necrosis Factor, a Tumor Necrosis Factor alpha, a Tumor necrosis factor beta, a Tumor necrosis factor receptor (TNFR), a VLA-4 protein, a VCAM-1 protein, a Vascular Endothelial Growth Factor (VEGEF), a Urokinase, a Mos, a Ras, a Raf, a Met; a p53, a Tat, a Fos, a Myc, a Jun, a Myb, a Rel, an estrogen receptor, a progesterone receptor, a testosterone receptor, an aldosterone receptor, an LDL receptor, a SCF/c-Kit, a CD40L/UCD40, a VLA-4/VCAM-1, an ICAM-1/LFA-1, a hyalurin/CD44.

Details for Patent 8,114,628

Applicant Tradename Biologic Ingredient Dosage Form BLA Approval Date Patent No. Expiredate
Eli Lilly And Company HUMULIN R U-100 insulin human Injection 018780 10/28/1982 ⤷  Try a Trial 2023-04-17
Eli Lilly And Company HUMULIN R U-500 insulin human Injection 018780 12/29/2015 ⤷  Try a Trial 2023-04-17
Eli Lilly And Company HUMULIN R U-100 insulin human Injection 018780 08/06/1998 ⤷  Try a Trial 2023-04-17
Eli Lilly And Company HUMULIN R U-500 insulin human Injection 018780 03/31/1994 ⤷  Try a Trial 2023-04-17
Eli Lilly And Company HUMULIN R U-100 insulin human Injection 018780 05/25/2018 ⤷  Try a Trial 2023-04-17
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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