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Title: Refactoring the Genetic Code for Increased Evolvability

ABSTRACT The standard genetic code is robust to mutations during transcription and translation. Point mutations are likely to be synonymous or to preserve the chemical properties of the original amino acid. Saturation mutagenesis experiments suggest that in some cases the best-performing mutant requires replacement of more than a single nucleotide within a codon. These replacements are essentially inaccessible to common error-based laboratory engineering techniques that alter a single nucleotide per mutation event, due to the extreme rarity of adjacent mutations. In this theoretical study, we suggest a radical reordering of the genetic code that maximizes the mutagenic potential of single nucleotide replacements. We explore several possible genetic codes that allow a greater degree of accessibility to the mutational landscape and may result in a hyperevolvable organism that could serve as an ideal platform for directed evolution experiments. We then conclude by evaluating the challenges of constructing such recoded organisms and their potential applications within the field of synthetic biology. IMPORTANCE The conservative nature of the genetic code prevents bioengineers from efficiently accessing the full mutational landscape of a gene via common error-prone methods. Here, we present two computational approaches to generate alternative genetic codes with increased accessibility. These new codesmore » allow mutational transitions to a larger pool of amino acids and with a greater extent of chemical differences, based on a single nucleotide replacement within the codon, thus increasing evolvability both at the single-gene and at the genome levels. Given the widespread use of these techniques for strain and protein improvement, along with more fundamental evolutionary biology questions, the use of recoded organisms that maximize evolvability should significantly improve the efficiency of directed evolution, library generation, and fitness maximization.« less
Authors:
; ; ; ;
Publication Date:
Grant/Contract Number:
SC008812
Type:
Published Article
Journal Name:
mBio (Online)
Additional Journal Information:
Journal Name: mBio (Online) Journal Volume: 8 Journal Issue: 6; Journal ID: ISSN 2150-7511
Publisher:
American Society for Microbiology
Sponsoring Org:
USDOE
Country of Publication:
United States
Language:
English
OSTI Identifier:
1437771

Pines, Gur, Winkler, James D., Pines, Assaf, Gill, Ryan T., and Lee, ed., Sang Yup. Refactoring the Genetic Code for Increased Evolvability. United States: N. p., Web. doi:10.1128/mBio.01654-17.
Pines, Gur, Winkler, James D., Pines, Assaf, Gill, Ryan T., & Lee, ed., Sang Yup. Refactoring the Genetic Code for Increased Evolvability. United States. doi:10.1128/mBio.01654-17.
Pines, Gur, Winkler, James D., Pines, Assaf, Gill, Ryan T., and Lee, ed., Sang Yup. 2017. "Refactoring the Genetic Code for Increased Evolvability". United States. doi:10.1128/mBio.01654-17.
@article{osti_1437771,
title = {Refactoring the Genetic Code for Increased Evolvability},
author = {Pines, Gur and Winkler, James D. and Pines, Assaf and Gill, Ryan T. and Lee, ed., Sang Yup},
abstractNote = {ABSTRACT The standard genetic code is robust to mutations during transcription and translation. Point mutations are likely to be synonymous or to preserve the chemical properties of the original amino acid. Saturation mutagenesis experiments suggest that in some cases the best-performing mutant requires replacement of more than a single nucleotide within a codon. These replacements are essentially inaccessible to common error-based laboratory engineering techniques that alter a single nucleotide per mutation event, due to the extreme rarity of adjacent mutations. In this theoretical study, we suggest a radical reordering of the genetic code that maximizes the mutagenic potential of single nucleotide replacements. We explore several possible genetic codes that allow a greater degree of accessibility to the mutational landscape and may result in a hyperevolvable organism that could serve as an ideal platform for directed evolution experiments. We then conclude by evaluating the challenges of constructing such recoded organisms and their potential applications within the field of synthetic biology. IMPORTANCE The conservative nature of the genetic code prevents bioengineers from efficiently accessing the full mutational landscape of a gene via common error-prone methods. Here, we present two computational approaches to generate alternative genetic codes with increased accessibility. These new codes allow mutational transitions to a larger pool of amino acids and with a greater extent of chemical differences, based on a single nucleotide replacement within the codon, thus increasing evolvability both at the single-gene and at the genome levels. Given the widespread use of these techniques for strain and protein improvement, along with more fundamental evolutionary biology questions, the use of recoded organisms that maximize evolvability should significantly improve the efficiency of directed evolution, library generation, and fitness maximization.},
doi = {10.1128/mBio.01654-17},
journal = {mBio (Online)},
number = 6,
volume = 8,
place = {United States},
year = {2017},
month = {11}
}

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