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

Xi, Zhiyan; Davis, Lloyd; Baxter, Kieran; Tynan, Ailish; Goutou, Angeliki; Greiss, Sebastian

doi:10.1093/nar/gkab1168

Cited by 21 publications

(6 citation statements)

References 50 publications

(76 reference statements)

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“…The activity regulation of Cre enzyme by GCE was previously proposed with success using engineered pyrrolysyl-tRNA synthetase/tRNA CUA (PylRS/tRNA CUA ) pairs to encode photocaged amino acids 21 , 22 . However, the efficiency of these developed PylRS/tRNA CUA pairs is low in yeast 23 .…”

Section: Resultsmentioning

confidence: 99%

Systematic dissection of key factors governing recombination outcomes by GCE-SCRaMbLE

Zhang

Gong

et al. 2022

Nat Commun

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With the completion of Sc2.0 chromosomes, synthetic chromosome rearrangement and modification by loxP-mediated evolution (SCRaMbLE) becomes more critical for in-depth investigation of fundamental biological questions and screening of industrially valuable characteristics. Further applications, however, are hindered due to the lack of facile and tight regulation of the SCRaMbLE process, and limited understanding of key factors that may affect the rearrangement outcomes. Here we propose an approach to precisely regulate SCRaMbLE recombination in a dose-dependent manner using genetic code expansion (GCE) technology with low basal activity. By systematically analyzing 1380 derived strains and six yeast pools subjected to GCE-SCRaMbLE, we find that Cre enzyme abundance, genome ploidy and chromosome conformation play key roles in recombination frequencies and determine the SCRaMbLE outcomes. With these insights, the GCE-SCRaMbLE system will serve as a powerful tool in the future exploitation and optimization of the Sc2.0-related technologies.

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Section: Resultsmentioning

confidence: 99%

Systematic dissection of key factors governing recombination outcomes by GCE-SCRaMbLE

Zhang

Gong

et al. 2022

Nat Commun

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“…The use of quadruplet codons provides greater flexibility, as it allows the incorporation of ncAAs without the need for large-scale genome modifications. This method also helps to reduce the potential cross-decoding effects of endogenous triplet codons [ 70 , 71 , 72 ]. Quadruplet codons have been primarily used in single-celled systems, such as the nematode Caenorhabditis elegans .…”

Section: Methods For Ncaas Incorporation Into Proteinsmentioning

confidence: 99%

Advances in Biosynthesis of Non-Canonical Amino Acids (ncAAs) and the Methods of ncAAs Incorporation into Proteins

Chen,

Xin,

Zhang

et al. 2023

Molecules

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The functional pool of canonical amino acids (cAAs) has been enriched through the emergence of non-canonical amino acids (ncAAs). NcAAs play a crucial role in the production of various pharmaceuticals. The biosynthesis of ncAAs has emerged as an alternative to traditional chemical synthesis due to its environmental friendliness and high efficiency. The breakthrough genetic code expansion (GCE) technique developed in recent years has allowed the incorporation of ncAAs into target proteins, giving them special functions and biological activities. The biosynthesis of ncAAs and their incorporation into target proteins within a single microbe has become an enticing application of such molecules. Based on that, in this study, we first review the biosynthesis methods for ncAAs and analyze the difficulties related to biosynthesis. We then summarize the GCE methods and analyze their advantages and disadvantages. Further, we review the application progress of ncAAs and anticipate the challenges and future development directions of ncAAs.

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“…Upon reaction with ONOO − , but not with other ROS, the moieties fall off and GFP turns on (Figure 4B). While none of the studies used their reporters in an in vivo system, genetic code expansion can be applied in animals [86].…”

Section: Superoxide Anion (Omentioning

confidence: 99%

Oxidative Stress in Age-Related Neurodegenerative Diseases: An Overview of Recent Tools and Findings

2023

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Reactive oxygen species (ROS) have been described to induce a broad range of redox-dependent signaling reactions in physiological conditions. Nevertheless, an excessive accumulation of ROS leads to oxidative stress, which was traditionally considered as detrimental for cells and organisms, due to the oxidative damage they cause to biomolecules. During ageing, elevated ROS levels result in the accumulation of damaged proteins, which may exhibit altered enzymatic function or physical properties (e.g., aggregation propensity). Emerging evidence also highlights the relationship between oxidative stress and age-related pathologies, such as protein misfolding-based neurodegenerative diseases (e.g., Parkinson’s (PD), Alzheimer’s (AD) and Huntington’s (HD) diseases). In this review we aim to introduce the role of oxidative stress in physiology and pathology and then focus on the state-of-the-art techniques available to detect and quantify ROS and oxidized proteins in live cells and in vivo, providing a guide to those aiming to characterize the role of oxidative stress in ageing and neurodegenerative diseases. Lastly, we discuss recently published data on the role of oxidative stress in neurological disorders.

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

Cited by 21 publications

References 50 publications

Systematic dissection of key factors governing recombination outcomes by GCE-SCRaMbLE

Systematic dissection of key factors governing recombination outcomes by GCE-SCRaMbLE

Advances in Biosynthesis of Non-Canonical Amino Acids (ncAAs) and the Methods of ncAAs Incorporation into Proteins

Oxidative Stress in Age-Related Neurodegenerative Diseases: An Overview of Recent Tools and Findings

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