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Compositions and methods for in vivo synthesis of non-natural polypeptides

A technology of unnatural and unnatural amino acids, applied in biochemical equipment and methods, using vectors to introduce foreign genetic material, polypeptides containing affinity tags, etc., can solve the problem of post-translational modification environment that is not sufficient to provide redox characteristics of unnatural polypeptides And other issues

Pending Publication Date: 2022-07-15
THE SCRIPPS RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, such expression systems are insufficient to provide a post-translational modification environment that can fully realize the redox properties of non-natural peptides and other post-translational modifications of synthetic non-natural peptides

Method used

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  • Compositions and methods for in vivo synthesis of non-natural polypeptides
  • Compositions and methods for in vivo synthesis of non-natural polypeptides
  • Compositions and methods for in vivo synthesis of non-natural polypeptides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach 1

[0262] Embodiment 1. A method of synthesizing a non-natural polypeptide, the method comprising:

[0263] a. providing at least one unnatural deoxyribonucleic acid (DNA) molecule comprising at least four unnatural base pairs;

[0264]b. Transcribe the at least one non-natural DNA molecule to obtain a messenger ribonucleic acid (mRNA) molecule comprising at least two non-natural codons;

[0265] c. Transcribe the at least one non-natural DNA molecule to obtain at least two transfer RNA (tRNA) molecules each comprising at least one non-natural anticodon, wherein the at least two non-natural base pairs in the corresponding DNA are in in a sequence context such that the non-natural codons of the mRNA molecules are complementary to the non-natural anti-codons of each of the tRNA molecules; and

[0266] d. synthesizing the non-natural polypeptide by translating the non-natural mRNA molecule using the at least two natural tRNA molecules, wherein each natural anticodon directs the sit...

Embodiment approach 11

[0267] Embodiment 1.1. A method of synthesizing a non-natural polypeptide, the method comprising:

[0268] a. providing at least one unnatural deoxyribonucleic acid (DNA) molecule comprising at least four unnatural base pairs;

[0269] b. Transcribe the at least one non-natural DNA molecule to obtain a messenger ribonucleic acid (mRNA) molecule comprising at least two non-natural codons;

[0270] c. Transcribe the at least one non-natural DNA molecule to obtain at least two transfer RNA (tRNA) molecules each comprising at least one non-natural anticodon, wherein the at least two non-natural base pairs in the corresponding DNA are in In a sequence context such that the non-natural codon of the mRNA molecule is complementary to the non-natural anti-codon of each of the tRNA molecules and at least one of the one or more other non-natural codons is complementary to the non-natural codon of each of the tRNA molecules. non-natural anticodon complementation of at least one of the ot...

Embodiment approach 2

[0272] Embodiment 2. A method of synthesizing a non-natural polypeptide, the method comprising:

[0273] a. providing at least one unnatural deoxyribonucleic acid (DNA) molecule comprising at least four unnatural base pairs, wherein the at least one unnatural DNA molecule encodes (i) comprises at least a first unnatural codon and a first two unnatural codon messenger ribonucleic acid (mRNA) molecules and (ii) at least a first transfer RNA (tRNA) molecule and a second transfer RNA molecule, the first tRNA molecule comprising a first unnatural anticodon and the The second tRNA molecule comprises a second unnatural anticodon, and the at least four unnatural base pairs in the at least one DNA molecule are in a sequence environment such that the first unnatural codon of the mRNA molecule and the first unnatural base pair two non-natural codons are complementary to the first non-natural anti-codon and the second non-natural anti-codon, respectively;

[0274] b. Transcribe the at le...

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Abstract

Disclosed herein are compositions, methods, and kits for cellular incorporation of non-natural amino acids into non-natural polypeptides. Also disclosed herein are compositions, methods, and kits for increasing the activity and yield of non-natural polypeptides synthesized from the cells.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS [0002] This application claims priority to US Provisional Application No. 62 / 913,664, filed October 10, 2019, and US Provisional Application No. 62 / 988,882, filed March 12, 2020, each of which is incorporated herein by reference in its entirety. [0003] sequence listing [0004] This application contains a Sequence Listing which has been electronically filed in ASCII format and is hereby incorporated by reference in its entirety. The ASCII copy created on October 6, 2010 is named "36271-809_601_SL.txt" and is 21 kilobytes in size. [0005] Statement Regarding Federally Sponsored Research [0006] This invention was made with US Government support under Grant No. GM118178 awarded by the National Institutes of Health. The United States Government has certain rights in this invention. [0007] incorporated by reference [0008] All publications, patents and patent applications mentioned in this specification are herein incorpor...

Claims

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Application Information

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IPC IPC(8): A61K31/7088C12N15/11C12N9/22
CPCC12N15/70C12Y601/01026C12Y207/07006C07K14/43595C07K2319/22C12P21/02C12N15/67C12N15/11C12N9/1247C12N9/93
Inventor F·E·罗姆斯伯格E·C·菲舍尔K·桥本A·W·费尔德曼V·T·迪恩Y·张
Owner THE SCRIPPS RES INST
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