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Using a quadruplet codon to expand the genetic code of an animal

Genetic code expansion in multicellular organisms is currently limited to the use of repurposed amber stop codons. Here, we introduce a system for the use of quadruplet codons to direct incorporation of non-canonical amino acids in vivo in an animal, the nematode worm Caenorhabditis elegans. We deve...

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Autores principales: Xi, Zhiyan, Davis, Lloyd, Baxter, Kieran, Tynan, Ailish, Goutou, Angeliki, Greiss, Sebastian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9122531/
https://www.ncbi.nlm.nih.gov/pubmed/34882769
http://dx.doi.org/10.1093/nar/gkab1168
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author Xi, Zhiyan
Davis, Lloyd
Baxter, Kieran
Tynan, Ailish
Goutou, Angeliki
Greiss, Sebastian
author_facet Xi, Zhiyan
Davis, Lloyd
Baxter, Kieran
Tynan, Ailish
Goutou, Angeliki
Greiss, Sebastian
author_sort Xi, Zhiyan
collection PubMed
description Genetic code expansion in multicellular organisms is currently limited to the use of repurposed amber stop codons. Here, we introduce a system for the use of quadruplet codons to direct incorporation of non-canonical amino acids in vivo in an animal, the nematode worm Caenorhabditis elegans. We develop hybrid pyrrolysyl tRNA variants to incorporate non-canonical amino acids in response to the quadruplet codon UAGA. We demonstrate the efficiency of the quadruplet decoding system by incorporating photocaged amino acids into two proteins widely used as genetic tools. We use photocaged lysine to express photocaged Cre recombinase for the optical control of gene expression and photocaged cysteine to express photo-activatable caspase for light inducible cell ablation. Our approach will facilitate the routine adoption of quadruplet decoding for genetic code expansion in eukaryotic cells and multicellular organisms.
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spelling pubmed-91225312022-05-23 Using a quadruplet codon to expand the genetic code of an animal Xi, Zhiyan Davis, Lloyd Baxter, Kieran Tynan, Ailish Goutou, Angeliki Greiss, Sebastian Nucleic Acids Res NAR Breakthrough Article Genetic code expansion in multicellular organisms is currently limited to the use of repurposed amber stop codons. Here, we introduce a system for the use of quadruplet codons to direct incorporation of non-canonical amino acids in vivo in an animal, the nematode worm Caenorhabditis elegans. We develop hybrid pyrrolysyl tRNA variants to incorporate non-canonical amino acids in response to the quadruplet codon UAGA. We demonstrate the efficiency of the quadruplet decoding system by incorporating photocaged amino acids into two proteins widely used as genetic tools. We use photocaged lysine to express photocaged Cre recombinase for the optical control of gene expression and photocaged cysteine to express photo-activatable caspase for light inducible cell ablation. Our approach will facilitate the routine adoption of quadruplet decoding for genetic code expansion in eukaryotic cells and multicellular organisms. Oxford University Press 2021-12-09 /pmc/articles/PMC9122531/ /pubmed/34882769 http://dx.doi.org/10.1093/nar/gkab1168 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle NAR Breakthrough Article
Xi, Zhiyan
Davis, Lloyd
Baxter, Kieran
Tynan, Ailish
Goutou, Angeliki
Greiss, Sebastian
Using a quadruplet codon to expand the genetic code of an animal
title Using a quadruplet codon to expand the genetic code of an animal
title_full Using a quadruplet codon to expand the genetic code of an animal
title_fullStr Using a quadruplet codon to expand the genetic code of an animal
title_full_unstemmed Using a quadruplet codon to expand the genetic code of an animal
title_short Using a quadruplet codon to expand the genetic code of an animal
title_sort using a quadruplet codon to expand the genetic code of an animal
topic NAR Breakthrough Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9122531/
https://www.ncbi.nlm.nih.gov/pubmed/34882769
http://dx.doi.org/10.1093/nar/gkab1168
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