Reagents and methods for replication, transcription, and translation in semi-synthetic organisms

Patent metadata

Field	Value
Publication number	US-11879145-B2
Application number	US-202016900154-A
Country	US
Kind code	B2
Filing date	Jun 12, 2020
Priority date	Jun 14, 2019
Publication date	Jan 23, 2024
Grant date	Jan 23, 2024

Abstract

Disclosed herein are compositions, methods, cells, engineered microorganisms, and kits for increasing the production of proteins or polypeptides comprising one or more unnatural amino acids. Further provided are compositions, cells, engineered microorganisms, and kits for increasing the retention of unnatural nucleic acids encoding the unnatural amino acids in an engineered cell, or semi-synthetic organism.

First claim

What is claimed is:

1. A nucleobase of the structure:

wherein:
each X is independently carbon or nitrogen;
R 2 is present when X is carbon and is independently hydrogen, alkyl, alkenyl, alkynyl, methoxy, methanethiol, methaneseleno, halogen, cyano, or azide group;
Y is sulfur, oxygen, or selenium; and
E is sulfur;
wherein the wavy line indicates a point of bonding to a ribosyl, deoxyribosyl, or dideoxyribosyl moiety, wherein the ribosyl, deoxyribosyl, or dideoxyribosyl moiety is optionally modified at the 2′ position, at the 3′ position, or at the 5′ position, wherein the ribosyl, deoxyribosyl, or dideoxyribosyl moiety is in free form, is connected to a mono-phosphate, diphosphate, triphosphate, α-thiotriphosphate, β-thiotriphosphate, or γ-thiotriphosphate group, or is included in a polynucleotide, optionally wherein the polynucleotide is an RNA, DNA, bicyclic nucleic acid, linked nucleic acid, peptide nucleic acid (PNA), locked nucleic acid (LNA), or a phosphorothioate-containing nucleic acid.

2. The nucleobase of claim 1 , wherein X is carbon and/or Y is sulfur.

3. The nucleobase of claim 1 , which has the structure

4. The nucleobase of claim 1 , which is bound to a complementary base-pairing nucleobase to form an unnatural base pair (UBP), wherein the complementary base-pairing nucleobase is selected from:

5. A double stranded oligonucleotide duplex wherein a first oligonucleotide strand comprises the nucleobase of claim 1 , and a second complementary oligonucleotide strand comprises a complementary base-pairing nucleobase in a complementary base-pairing site thereof.

6. The double stranded oligonucleotide duplex of claim 5 , wherein the first oligonucleotide strand comprises

and the second strand comprises a complementary base pairing nucleobase selected from:

in a complementary base-pairing site thereof.

7. A DNA comprising a nucleobase having the structure

and a complementary base-pairing nucleobase having the structure

or a nucleobase having the structure

and a complementary base-pairing nucleobase having the structure

Assignees

• Scripps Research Inst

Inventors

• Romesberg Floyd E
• Dien Vivian T
• Feldman Aaron W
• Karadeema Rebekah J
• Li Lingjun
• Ledbetter Michael P
• Zhou Anne Xiaozhou

Classifications

• C07H21/00

  Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids · CPC title

• C07H19/00 Primary

  Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides ; Anhydro-derivatives thereof · CPC title

• C12N15/11 Primary

  DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity} · CPC title

• C12P19/34 Primary

  Polynucleotides, e.g. nucleic acids, oligoribonucleotides · CPC title

• C12P21/02

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title