Genetic encryption
View Patent ↗The present invention relates to a method to engineer either the genome of a genetically modified organism, other bioengineered reagent, or in vitro translation system for protein synthesis from specific protein-coding genes so that the protein-coding genes so engineered can only produce proteins with an intended structure when translated within the context of that specifically engineered GMO or in vitro translation system. It also relates to nucleic acids for use in such GMOs or translation systems.
1. A genetically engineered prokaryotic cell comprising a genome that comprises, compared to a corresponding wild-type prokaryotic cell having wild-type genetic code:
(a) substitutions of all occurrences of at least two codons, in one or more protein-encoding sequences in the genome, with substituted codons that code for different amino acids according to the wild-type genetic code, and
(b) alterations to tRNA genes such that the altered tRNA comprise anticodons cognate to unsubstituted codons but carry amino acids that the unsubstituted codons code for according to the wild-type genetic code,
thereby the one or more protein-encoding sequence in the genetically engineered cell encodes the same amino acid sequence as the one or more protein-encoding sequence in the wild-type cell.
2. The cell of claim 1 wherein all occurrences of a first codon are substituted with a second codon and all occurrences of the second codon are substituted with the first codon.
3. The cell of claim 1 wherein all occurrences of codons coding for a first amino acid according to the wild-type genetic code are substituted with codons that code for a second amino acid according to the wild-type genetic code.
4. The cell of claim 1 , wherein the alterations of the tRNA genes comprise substituting nucleotides at positions 1-26 and 44-76 of the tRNA with nucleotides at positions 1-26 and 44-76 from the tRNAs that carry amino acids that the unsubstituted codons code for according to the wild-type genetic code.
5. The cell of claim 1 , wherein the alterations of the tRNA genes comprise substituting nucleotides at positions 1 through 26, 27, 28, 29, 30, 31, 32 or 33 and 37, 38, 39, 40, 41, 42, 43 or 44 through 76 of the tRNA with nucleotides at positions 1 through 26, 27, 28, 29, 30, 31, 32 or 33 and 37, 38, 39, 40, 41, 42, 43 or 44 through 76 from the tRNAs that carry amino acids that the unsubstituted codons code for according to the wild-type genetic code.
6. The cell of claim 1 , wherein codons coding for serine are substituted with codons coding for leucine or alanine, both according to the wild-type genetic code.
7. The cell of claim 1 , wherein codons coding for leucine are substituted with codons coding for serine or alanine, both according to the wild-type genetic code.
8. The cell of claim 1 , wherein codons coding for alanine are substituted with codons coding for serine or leucine, both according to the wild-type genetic code.
9. The cell of claim 1 , wherein codons coding for serine are substituted with codons coding for leucine or alanine, codons coding for leucine are substituted with codons coding for alanine or serine, and codons coding for alanine are substituted with codons coding for serine or leucine, all according to the wild-type genetic code.
10. The cell of claim 1 , wherein the codon substitutions comprise one or more provided in Table 1.
11. An in vitro translation system derived from the cell of claim 1 .
12. A method for preparing a genetically engineered prokaryotic cell, comprising:
(i) preparing a genome that comprises, compared to a corresponding wild-type prokaryotic cell having wild-type genetic code:
(a) substitutions of all occurrences of at least two codons, in one or more protein-encoding sequences in the genome, with substituted codons that code for different amino acids according to the wild-type genetic code, and
(b) alterations to tRNA genes corresponding to the substituted codons such that the altered tRNA carry amino acids that the unsubstituted codons code for according to the wild-type genetic code,
thereby the one or more protein-encoding sequence in the genetically engineered cell encodes the same amino acid sequence as the one or more protein-encoding sequence in the wild-type cell, and
(ii) transformed the genome into a prokaryotic cell that does not contain genomic material.
13. The method of claim 12 , wherein the alterations of the tRNA genes comprise substituting nucleotides at positions 1-26 and 44-76 of the tRNA with nucleotides at positions 1-26 and 44-76 from the tRNAs that carry amino acids that the unsubstituted codons code for according to the wild-type genetic code.
14. The method of claim 12 , wherein the alterations of the tRNA genes comprise substituting nucleotides at positions 1 through 26, 27, 28, 29, 30, 31, 32 or 33 and 37, 38, 39, 40, 41, 42, 43 or 44 through 76 of the tRNA with nucleotides at positions 1 through 26, 27, 28, 29, 30, 31, 32 or 33 and 37, 38, 39, 40, 41, 42, 43 or 44 through 76 from the tRNAs that carry amino acids that the unsubstituted codons code for according to the wild-type genetic code.
15. The method of claim 12 , wherein all occurrences of a first codon are substituted with a second codon and all occurrences of the second codon are substituted with the first codon.
16. The method of claim 12 , wherein all occurrences of codons coding for a first amino acid according to the wild-type genetic code are substituted with codons that code for a second amino acid according to the wild-type.
17. The method of claim 12 , wherein codons coding for serine are substituted with codons coding for leucine or alanine, both according to the wild-type genetic code.
18. The method of claim 12 , wherein codons coding for leucine are substituted with codons coding for serine or alanine, both according to the wild-type genetic code.
19. The method of claim 12 , wherein codons coding for alanine are substituted with codons coding for serine or leucine, both according to the wild-type genetic code.
20. The method of claim 12 , wherein codons coding for serine are substituted with codons coding for leucine or alanine, codons coding for leucine are substituted with codons coding for alanine or serine, and codons coding for alanine are substituted with codons coding for serine or leucine, all according to the wild-type genetic code.
21. The method of claim 12 , wherein the codon substitutions comprise one or more provided in Table 1.