Ligand-responsive upregulation of 3′ CITE-mediated translation in a wheat germ cell-free expression system

Ogawa, Atsushi; Murashige, Yuta; Tabuchi, Junichiro; Omatsu, Taiki

doi:10.1039/c6mb00748a

Cited by 14 publications

(24 citation statements)

References 38 publications

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“…Last but not least, the diversity of cis-acting translation elements identified in plant viruses may contribute to the design of tools for synthetic biology (Ogawa et al, 2017), and in vectors for the overexpression of proteins in biofactory cell-free systems, cell cultures, or whole plants (Fan et al, 2012), or, perhaps, in other organisms used for industrial overexpression of proteins, if mechanisms employed by plant viruses are universal or at least conserved in the species of interest.…”

Section: Perspectivesmentioning

confidence: 99%

Non-canonical Translation in Plant RNA Viruses

et al. 2017

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Viral protein synthesis is completely dependent upon the host cell's translational machinery. Canonical translation of host mRNAs depends on structural elements such as the 5′ cap structure and/or the 3′ poly(A) tail of the mRNAs. Although many viral mRNAs are devoid of one or both of these structures, they can still translate efficiently using non-canonical mechanisms. Here, we review the tools utilized by positive-sense single-stranded (+ss) RNA plant viruses to initiate non-canonical translation, focusing on cis-acting sequences present in viral mRNAs. We highlight how these elements may interact with host translation factors and speculate on their contribution for achieving translational control. We also describe other translation strategies used by plant viruses to optimize the usage of the coding capacity of their very compact genomes, including leaky scanning initiation, ribosomal frameshifting and stop-codon readthrough. Finally, future research perspectives on the unusual translational strategies of +ssRNA viruses are discussed, including parallelisms between viral and host mRNAs mechanisms of translation, particularly for host mRNAs which are translated under stress conditions.

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Section: Perspectivesmentioning

confidence: 99%

Non-canonical Translation in Plant RNA Viruses

et al. 2017

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“…Unfortunately all results for these constructs were generated in cell-free translation system using wheat germ extract, making their functionality in human cells an open question. In this vein, Ogawa et al also report aptamer-mediated regulation of 3 cap-independent translation elements which mediate alternative initiation mechanisms in plant viruses but are unsuitable for use in mammals [94]. However, IRES-mediated initiation is employed both by human-tropic viruses and human cells, suggesting that this strategy may be transferable [95].…”

Section: Riboswitches Controlling Alternative Initiation Mechanismsmentioning

confidence: 99%

Riboswitches for Controlled Expression of Therapeutic Transgenes Delivered by Adeno-Associated Viral Vectors

Tickner

Farzan

2021

Pharmaceuticals

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Vectors developed from adeno-associated virus (AAV) are powerful tools for in vivo transgene delivery in both humans and animal models, and several AAV-delivered gene therapies are currently approved for clinical use. However, AAV-mediated gene therapy still faces several challenges, including limited vector packaging capacity and the need for a safe, effective method for controlling transgene expression during and after delivery. Riboswitches, RNA elements which control gene expression in response to ligand binding, are attractive candidates for regulating expression of AAV-delivered transgene therapeutics because of their small genomic footprints and non-immunogenicity compared to protein-based expression control systems. In addition, the ligand-sensing aptamer domains of many riboswitches can be exchanged in a modular fashion to allow regulation by a variety of small molecules, proteins, and oligonucleotides. Riboswitches have been used to regulate AAV-delivered transgene therapeutics in animal models, and recently developed screening and selection methods allow rapid isolation of riboswitches with novel ligands and improved performance in mammalian cells. This review discusses the advantages of riboswitches in the context of AAV-delivered gene therapy, the subsets of riboswitch mechanisms which have been shown to function in human cells and animal models, recent progress in riboswitch isolation and optimization, and several examples of AAV-delivered therapeutic systems which might be improved by riboswitch regulation.

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“…36 The sequences of these synthetic nucleic acids are described in Supporting Information. The sequence of a synthetic plasmid, pEU-nLuc-Venus encoding the NanoLuc gene, 37 has been reported elesewhere 21 and that of another synthetic plasmid, pTheo5-PSIV-IRES encoding the PSIV (Plautia stali intestine virus) IRES, 38 is described in Supporting Information.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

In Vitro Selection of RNA Aptamers Binding to Nanosized DNA for Constructing Artificial Riboswitches

Ogawa

Itoh

2020

ACS Synth. Biol.

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We here designed an in vitro selection scheme for obtaining an aptamer with which to rationally construct an artificial riboswitch as its component part. In fact, a nanosized DNA-binding aptamer obtained through this scheme allowed us to easily and successfully create eukaryotic riboswitches that upregulate internal ribosome entry site-mediated translation in response to the ligand (nanosized DNA) in wheat germ extract, a eukaryotic cell-free expression system. The induction ratio of the best riboswitch ligand-dose-dependently increased to 21 at 300 μM ligand. This switching efficiency is much higher than that of the same type of riboswitch with a widely used theophylline-binding aptamer, which was in vitro selected without considering its utility for constructing riboswitches. The selection scheme described here would facilitate obtaining various ligand/aptamer pairs suitable for constructing artificial riboswitches, which could serve as elements of synthetic gene circuits in synthetic biology.

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Ligand-responsive upregulation of 3′ CITE-mediated translation in a wheat germ cell-free expression system

Cited by 14 publications

References 38 publications

Non-canonical Translation in Plant RNA Viruses

Non-canonical Translation in Plant RNA Viruses

Riboswitches for Controlled Expression of Therapeutic Transgenes Delivered by Adeno-Associated Viral Vectors

In Vitro Selection of RNA Aptamers Binding to Nanosized DNA for Constructing Artificial Riboswitches

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