Extensive breaking of genetic code degeneracy with non-canonical amino acids
Abstract Genetic code expansion (GCE) offers many exciting opportunities for the creation of synthetic organisms and for drug discovery methods that utilize in vitro translation. One type of GCE, sense codon reassignment (SCR), focuses on breaking the degeneracy of the 61 sense codons which encode f...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-08-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-40529-x |
| _version_ | 1797558391172759552 |
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| author | Clinton A. L. McFeely Bipasana Shakya Chelsea A. Makovsky Aidan K. Haney T. Ashton Cropp Matthew C. T. Hartman |
| author_facet | Clinton A. L. McFeely Bipasana Shakya Chelsea A. Makovsky Aidan K. Haney T. Ashton Cropp Matthew C. T. Hartman |
| author_sort | Clinton A. L. McFeely |
| collection | DOAJ |
| description | Abstract Genetic code expansion (GCE) offers many exciting opportunities for the creation of synthetic organisms and for drug discovery methods that utilize in vitro translation. One type of GCE, sense codon reassignment (SCR), focuses on breaking the degeneracy of the 61 sense codons which encode for only 20 amino acids. SCR has great potential for genetic code expansion, but extensive SCR is limited by the post-transcriptional modifications on tRNAs and wobble reading of these tRNAs by the ribosome. To better understand codon-tRNA pairing, here we develop an assay to evaluate the ability of aminoacyl-tRNAs to compete with each other for a given codon. We then show that hyperaccurate ribosome mutants demonstrate reduced wobble reading, and when paired with unmodified tRNAs lead to extensive and predictable SCR. Together, we encode seven distinct amino acids across nine codons spanning just two codon boxes, thereby demonstrating that the genetic code hosts far more re-assignable space than previously expected, opening the door to extensive genetic code engineering. |
| first_indexed | 2024-03-10T17:30:48Z |
| format | Article |
| id | doaj.art-8fac6733bce441ee8239a9a3fc923092 |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-10T17:30:48Z |
| publishDate | 2023-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-8fac6733bce441ee8239a9a3fc9230922023-11-20T10:02:00ZengNature PortfolioNature Communications2041-17232023-08-0114111310.1038/s41467-023-40529-xExtensive breaking of genetic code degeneracy with non-canonical amino acidsClinton A. L. McFeely0Bipasana Shakya1Chelsea A. Makovsky2Aidan K. Haney3T. Ashton Cropp4Matthew C. T. Hartman5Department of Chemistry, Virginia Commonwealth UniversityDepartment of Chemistry, Virginia Commonwealth UniversityDepartment of Chemistry, Virginia Commonwealth UniversityDepartment of Chemistry, Virginia Commonwealth UniversityDepartment of Chemistry, Virginia Commonwealth UniversityDepartment of Chemistry, Virginia Commonwealth UniversityAbstract Genetic code expansion (GCE) offers many exciting opportunities for the creation of synthetic organisms and for drug discovery methods that utilize in vitro translation. One type of GCE, sense codon reassignment (SCR), focuses on breaking the degeneracy of the 61 sense codons which encode for only 20 amino acids. SCR has great potential for genetic code expansion, but extensive SCR is limited by the post-transcriptional modifications on tRNAs and wobble reading of these tRNAs by the ribosome. To better understand codon-tRNA pairing, here we develop an assay to evaluate the ability of aminoacyl-tRNAs to compete with each other for a given codon. We then show that hyperaccurate ribosome mutants demonstrate reduced wobble reading, and when paired with unmodified tRNAs lead to extensive and predictable SCR. Together, we encode seven distinct amino acids across nine codons spanning just two codon boxes, thereby demonstrating that the genetic code hosts far more re-assignable space than previously expected, opening the door to extensive genetic code engineering.https://doi.org/10.1038/s41467-023-40529-x |
| spellingShingle | Clinton A. L. McFeely Bipasana Shakya Chelsea A. Makovsky Aidan K. Haney T. Ashton Cropp Matthew C. T. Hartman Extensive breaking of genetic code degeneracy with non-canonical amino acids Nature Communications |
| title | Extensive breaking of genetic code degeneracy with non-canonical amino acids |
| title_full | Extensive breaking of genetic code degeneracy with non-canonical amino acids |
| title_fullStr | Extensive breaking of genetic code degeneracy with non-canonical amino acids |
| title_full_unstemmed | Extensive breaking of genetic code degeneracy with non-canonical amino acids |
| title_short | Extensive breaking of genetic code degeneracy with non-canonical amino acids |
| title_sort | extensive breaking of genetic code degeneracy with non canonical amino acids |
| url | https://doi.org/10.1038/s41467-023-40529-x |
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