A protocol to construct RNA-protein devices for photochemical translational regulation of synthetic mRNAs in mammalian cells

Here, we describe a protocol for the translational regulation of transfected messenger RNAs (mRNAs) using light in mammalian cells. We detail the steps for photocaged ligand synthesis, template DNA preparation, and mRNA synthesis. We describe steps for mRNA transfection, treatment of cells with a ph...

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Detalles Bibliográficos
Autores principales: Nakanishi, Hideyuki, Yoshii, Tatsuyuki, Tsukiji, Shinya, Saito, Hirohide
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9189627/
https://www.ncbi.nlm.nih.gov/pubmed/35707682
http://dx.doi.org/10.1016/j.xpro.2022.101451
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author Nakanishi, Hideyuki
Yoshii, Tatsuyuki
Tsukiji, Shinya
Saito, Hirohide
author_facet Nakanishi, Hideyuki
Yoshii, Tatsuyuki
Tsukiji, Shinya
Saito, Hirohide
author_sort Nakanishi, Hideyuki
collection PubMed
description Here, we describe a protocol for the translational regulation of transfected messenger RNAs (mRNAs) using light in mammalian cells. We detail the steps for photocaged ligand synthesis, template DNA preparation, and mRNA synthesis. We describe steps for mRNA transfection, treatment of cells with a photocaged ligand followed by light irradiation, and analysis of the transgene expression. The protocol enables spatiotemporally regulated transgene expression without the risk of insertional mutagenesis. For complete details on the use and execution of this protocol, please refer to Nakanishi et al. (2021).
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spelling pubmed-91896272022-06-14 A protocol to construct RNA-protein devices for photochemical translational regulation of synthetic mRNAs in mammalian cells Nakanishi, Hideyuki Yoshii, Tatsuyuki Tsukiji, Shinya Saito, Hirohide STAR Protoc Protocol Here, we describe a protocol for the translational regulation of transfected messenger RNAs (mRNAs) using light in mammalian cells. We detail the steps for photocaged ligand synthesis, template DNA preparation, and mRNA synthesis. We describe steps for mRNA transfection, treatment of cells with a photocaged ligand followed by light irradiation, and analysis of the transgene expression. The protocol enables spatiotemporally regulated transgene expression without the risk of insertional mutagenesis. For complete details on the use and execution of this protocol, please refer to Nakanishi et al. (2021). Elsevier 2022-06-10 /pmc/articles/PMC9189627/ /pubmed/35707682 http://dx.doi.org/10.1016/j.xpro.2022.101451 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Nakanishi, Hideyuki
Yoshii, Tatsuyuki
Tsukiji, Shinya
Saito, Hirohide
A protocol to construct RNA-protein devices for photochemical translational regulation of synthetic mRNAs in mammalian cells
title A protocol to construct RNA-protein devices for photochemical translational regulation of synthetic mRNAs in mammalian cells
title_full A protocol to construct RNA-protein devices for photochemical translational regulation of synthetic mRNAs in mammalian cells
title_fullStr A protocol to construct RNA-protein devices for photochemical translational regulation of synthetic mRNAs in mammalian cells
title_full_unstemmed A protocol to construct RNA-protein devices for photochemical translational regulation of synthetic mRNAs in mammalian cells
title_short A protocol to construct RNA-protein devices for photochemical translational regulation of synthetic mRNAs in mammalian cells
title_sort protocol to construct rna-protein devices for photochemical translational regulation of synthetic mrnas in mammalian cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9189627/
https://www.ncbi.nlm.nih.gov/pubmed/35707682
http://dx.doi.org/10.1016/j.xpro.2022.101451
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