Directed Evolution: Methodologies and Applications

Wang, Yajie; Xue, Pu; Cao, Mingfeng; Yu, Tianhao; Lane, Stephan; Zhao, Huimin

doi:10.1021/acs.chemrev.1c00260

Cited by 305 publications

(269 citation statements)

References 650 publications

(1,229 reference statements)

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“…The success of this viral-engineering strategy (1) defined guidelines for UAA incorporation and viral generation that could be broadly expanded to other RNA viruses in this study (2) provided a potential solution to the immediate quandary of vaccination by harnessing EV71-NAEK to elicit strong immune responses that were adjustable via artificial NAEK administration. For UAA-mediated viral engineering, although genetic codon expansion technology has been employed to modify virus-like particles 29,30 , engineer viral vectors for gene therapy 31,32 , and probe the biology of viruses 33,34 , UAAincorporated sites are generally selected by random mutations within targeted viral proteins, a traditional screening method for protein engineering 35 , and the lack of defined rules considering both protein modification and virus assembly. Moreover, engineered viruses developed in previous studies were always packaged in HEK293T cells with transiently or stably expressed PylRS/tRNA pairs 18 , which is appropriate for generation of common viral vectors (e.g., adeno-associated viruses [AAVs], lentiviruses) but not for most viruses' propagation, especially in vaccine development 36 .…”

Section: Discussionmentioning

confidence: 99%

Engineering RNA viruses with unnatural amino acid to evoke adjustable immune response in mice

Zheng

Wang

et al. 2021

Preprint

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Ribonucleic acid (RNA) viruses pose heavy burdens on public-health systems. Synthetic biology holds great potential for artificially controlling their replication, a strategy that could be used to attenuate infectious viruses but is still in the exploratory stage. Herein, we used the genetic-code expansion technique to convert Enterovirus 71 (EV71), a model of RNA virus, into a controllable EV71 strain carrying the unnatural amino acid (UAA) Nε-2-azidoethyloxycarbonyl-L-lysine (NAEK), which we termed an EV71-NAEK virus. EV71-NAEK could recapitulate an authentic NAEK time- and dose-dependent infection in vitro and in vivo, which could serve as a novel method to manipulate virulent viruses in conventional laboratories. We further validated the prophylactic effect of EV71-NAEK in two mouse models. In susceptible parent mice, vaccination with EV71-NAEK elicited a strong immune response and potentially protected their neonatal offspring from lethal challenge similar to that of commercial vaccines. Meanwhile, in transgenic mice harboring a PylRS-tRNAPyl CUA pair, substantial elements of genetic-code expansion technology, EV71-NAEK evoked an adjustable neutralizing-antibody response in a strictly external NAEK dose-dependent manner. These findings suggested that EV71-NAEK could be the basis of a feasible immunization program for populations with different levels of immunity. Moreover, we expanded the strategy to generate controllable coxsackieviruses and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) for conceptual verification. In combination, these results could underlie a competent strategy for attenuating viruses and priming the immune system via artificial control, which might be a promising direction for the development of amenable vaccine candidates and be broadly applied to other RNA viruses.

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Section: Discussionmentioning

confidence: 99%

Engineering RNA viruses with unnatural amino acid to evoke adjustable immune response in mice

Zheng

Wang

et al. 2021

Preprint

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“…Microorganisms cannot produce enough targeted product because of enzymes that have limited turnover or poor expression ( Song et al, 2017 ; Song et al, 2018 ; Lian et al, 2018 ). Therefore, protein engineering, particularly directed evolution, has become one of the most powerful and widespread tools for engineering improved or novel functions in enzymes ( Wu et al, 2013 ; Wang et al, 2021 ). Researchers employed protein engineering of 4CL and STS in E. coli for higher and more efficient resveratrol production ( Becker et al, 2003 ; Zhang et al, 2015 ).…”

Section: Enzyme Engineeringmentioning

confidence: 99%

Synthetic Biology-Driven Microbial Production of Resveratrol: Advances and Perspectives

Feng

Chen

et al. 2022

Front. Bioeng. Biotechnol.

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Resveratrol, a bioactive natural product found in many plants, is a secondary metabolite and has attracted much attention in the medicine and health care products fields due to its remarkable biological activities including anti-cancer, anti-oxidation, anti-aging, anti-inflammation, neuroprotection and anti-glycation. However, traditional chemical synthesis and plant extraction methods are impractical for industrial resveratrol production because of low yield, toxic chemical solvents and environmental pollution during the production process. Recently, the biosynthesis of resveratrol by constructing microbial cell factories has attracted much attention, because it provides a safe and efficient route for the resveratrol production. This review discusses the physiological functions and market applications of resveratrol. In addition, recent significant biotechnology advances in resveratrol biosynthesis are systematically summarized. Furthermore, we discuss the current challenges and future prospects for strain development for large-scale resveratrol production at an industrial level.

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“…This complex genetic expression can be achieved by using genetic circuits (Kim et al, 2018). Genetic circuits are an assembly of transgenes, referred to as biological parts, and encoding proteins or in some cases untranslated RNA that allows specific input signals to be detected and interpreted to control the expression of an output RNA or protein (Wang, Xue, et al, 2021). The circuit constructed may be simple or complex, but the goal is to engineer a virus that can detect whether it has infected a normal cell or a tumor cell and subsequently respond by replicating in and killing tumor cells while not replicating in normal cells (Tripodi et al, 2021).…”

Section: Virotherapymentioning

confidence: 99%

Smart Oncolytic Adenovirotherapy to Induce Killing of Cancer Cells and Elicit Antitumor Immunity

Enekegho¹,

Stuart²

2022

Eureka

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Cancer is one of the leading causes of death in the world, accounting for over 30% of all deaths in Canada. Various chemotherapy and therapeutic agents are currently in practice to help combat and treat cancerous growths and to lead to cancer remission. Virotherapy is an emerging treatment that uses biotechnology to convert viruses into therapeutic agents for the treatment of specific types of cancer. This process reprograms viruses to become oncolytic and target tumor cells in the body for lysis. It also uses these viruses to recruit inflammatory and vaccination responses by the immune system to help kill surrounding tumor cells while also establishing a long immune memory to help in the case of later infections. Adenoviruses are a group of viruses that infect the membranes of the respiratory tract, eyes, intestines, urinary tract, and nervous system of humans and causing fever as well as many cold symptoms. It is also a commonly used oncolytic virus and has been demonstrated in recent studies to be a great potential tool for eliciting appropriate inflammatory responses from the immune system to kill cancer cells and inducing cell-mediated immunity to prevent against later re-infection by the specific cancer type. Advances to this virotherapy has progressed towards overcoming tumor-mediated immunosuppression, which usually allows cancerous cells to evade the immune system and escape cell destruction, especially when combined with other therapy treatments. (Goradel et al., 2019). This review will focus on the mechanism as to how engineered modified viruses stimulate the immune system for cell killing and cell-mediated immunity. There will also be an examination of several research papers with some evidence to understand the synergy being oncolytic adenovirotherapy and the immune system function to kill cancer cells. Some disadvantages and issues with using this form of therapeutic treatment will also be presented, as well as some present and future research operating to fix these issues as well as increase the overall efficacy of this cancer treatment oncolytic adenovirotherapy.

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Directed Evolution: Methodologies and Applications

Cited by 305 publications

References 650 publications

Engineering RNA viruses with unnatural amino acid to evoke adjustable immune response in mice

Engineering RNA viruses with unnatural amino acid to evoke adjustable immune response in mice

Synthetic Biology-Driven Microbial Production of Resveratrol: Advances and Perspectives

Smart Oncolytic Adenovirotherapy to Induce Killing of Cancer Cells and Elicit Antitumor Immunity

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