INCORPORATION OF UNNATURAL NUCLEOTIDES AND METHODS THEREOF

• US 20200131555A1
• Filed: 07/10/2018
• Published: 04/30/2020
• Est. Priority Date: 07/11/2017
• Status: Abandoned Application

• Current Assignee: Synthorx Inc. (Sanofi ), The Scripps Research Institute
• Original Assignee: Synthorx Inc. (Sanofi ), The Scripps Research Institute
• Inventors: PTACIN, Jerod, CAFFARO, Carolina, AERNI, Hans, ZHANG, Yorke, FISCHER, Emil C., FELDMAN, Aaron W., DIEN, Vivian T., ROMESBERG, Floyd E.
• Application Number:
  US16/629,255
• Publication Number:
  US 20200131555A1
• Time in Patent Office: Days
• CPC Class Codes: C12N 15/11 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 Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/90 Stable introduction of foreign DNA into chromosome
• CPC Class Codes: C12N 9/93 Ligases (6)
• CPC Class Codes: C12P 21/02 having a known sequence of two or more amino acids, e.g. glutathione
• CPC Class Codes: C12Y 601/01026 Pyrrolysine-tRNAPyl ligase (6.1.1.26)

Abstract

Disclosed herein are methods, compositions and kits for the synthesis of proteins which comprises unnatural amino acids that utilize a mutant tRNA.

Claims

1-23. -23. (canceled)

24. An E. coli cell comprising:

(a) a nucleoside triphosphate transporter from Phaeodactylum tricornutum;

(b) an mRNA comprising an unnatural base comprising a structure selected from

25. The E. coli cell of claim 24, wherein the cell further comprises an oligonucleotide encoding the mRNA.

26. The E. coli cell of claim 24, wherein the cell further comprises an oligonucleotide encoding the tRNA.

27. The E. coli cell of claim 24, wherein the cell further comprises an oligonucleotide encoding the aminoacyl tRNA synthetase.

28. The E. coli cell of claim 24, wherein the cell further comprises a single oligonucleotide encoding the mRNA and the tRNA.

29. The E. coli cell of claim 24, wherein the cell further comprises a single oligonucleotide encoding the mRNA, the tRNA, and the aminoacyl tRNA synthetase.

30. The E. coli cell of claim 24, wherein the mRNA comprises one or more codons selected from AXC and GXC, wherein X is the unnatural base.

31. The E. coli cell of claim 30, wherein the mRNA comprises the codon AXC.

32. The E. coli cell of claim 31, wherein the unnatural base comprises the structure

33. The E. coli cell of claim 31, wherein the unnatural base comprises the structure

34. The E. coli cell of claim 30, wherein the mRNA comprises the codon GXC.

35. The E. coli cell of claim 34, wherein the unnatural base comprises the structure

36. The E. coli cell of claim 34, wherein the unnatural base comprises the structure

37. The E. coli cell of claim 24, wherein the aminoacyl tRNA synthetase is derived from Methanosarcina barkeri.

38. The E. coli cell of claim 36, wherein the mRNA comprises an unnatural base comprising the structure

39. The E. coli cell of claim 37, wherein the mRNA comprises one or more codons selected from AXC and GXC, wherein X is the unnatural base.

40. The E. coli cell of claim 24, wherein the aminoacyl tRNA synthetase is derived from Methanococcus jannaschii.

41. The E. coli cell of claim 39, wherein the mRNA comprises an unnatural base comprising the structure

42. The E. coli cell of claim 40, wherein the mRNA comprises one or more codons selected from AXC and GXC, wherein X is the unnatural base.

43. The E. coli cell of claim 24, wherein the tRNA is derived from an organism selected from the group consisting of Methanosarcina barkeri and Methanococcus jannaschii.