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Synthetic mRNA devices that detect endogenous proteins and distinguish mammalian cells

Synthetic biology has great potential for future therapeutic applications including autonomous cell programming through the detection of protein signals and the production of desired outputs. Synthetic RNA devices are promising for this purpose. However, the number of available devices is limited du...

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Autores principales: Kawasaki, Shunsuke, Fujita, Yoshihiko, Nagaike, Takashi, Tomita, Kozo, Saito, Hirohide
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5499560/
https://www.ncbi.nlm.nih.gov/pubmed/28525643
http://dx.doi.org/10.1093/nar/gkx298
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author Kawasaki, Shunsuke
Fujita, Yoshihiko
Nagaike, Takashi
Tomita, Kozo
Saito, Hirohide
author_facet Kawasaki, Shunsuke
Fujita, Yoshihiko
Nagaike, Takashi
Tomita, Kozo
Saito, Hirohide
author_sort Kawasaki, Shunsuke
collection PubMed
description Synthetic biology has great potential for future therapeutic applications including autonomous cell programming through the detection of protein signals and the production of desired outputs. Synthetic RNA devices are promising for this purpose. However, the number of available devices is limited due to the difficulty in the detection of endogenous proteins within a cell. Here, we show a strategy to construct synthetic mRNA devices that detect endogenous proteins in living cells, control translation and distinguish cell types. We engineered protein-binding aptamers that have increased stability in the secondary structures of their active conformation. The designed devices can efficiently respond to target proteins including human LIN28A and U1A proteins, while the original aptamers failed to do so. Moreover, mRNA delivery of an LIN28A-responsive device into human induced pluripotent stem cells (hiPSCs) revealed that we can distinguish living hiPSCs and differentiated cells by quantifying endogenous LIN28A protein expression level. Thus, our endogenous protein-driven RNA devices determine live-cell states and program mammalian cells based on intracellular protein information.
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spelling pubmed-54995602017-07-10 Synthetic mRNA devices that detect endogenous proteins and distinguish mammalian cells Kawasaki, Shunsuke Fujita, Yoshihiko Nagaike, Takashi Tomita, Kozo Saito, Hirohide Nucleic Acids Res Methods Online Synthetic biology has great potential for future therapeutic applications including autonomous cell programming through the detection of protein signals and the production of desired outputs. Synthetic RNA devices are promising for this purpose. However, the number of available devices is limited due to the difficulty in the detection of endogenous proteins within a cell. Here, we show a strategy to construct synthetic mRNA devices that detect endogenous proteins in living cells, control translation and distinguish cell types. We engineered protein-binding aptamers that have increased stability in the secondary structures of their active conformation. The designed devices can efficiently respond to target proteins including human LIN28A and U1A proteins, while the original aptamers failed to do so. Moreover, mRNA delivery of an LIN28A-responsive device into human induced pluripotent stem cells (hiPSCs) revealed that we can distinguish living hiPSCs and differentiated cells by quantifying endogenous LIN28A protein expression level. Thus, our endogenous protein-driven RNA devices determine live-cell states and program mammalian cells based on intracellular protein information. Oxford University Press 2017-07-07 2017-05-19 /pmc/articles/PMC5499560/ /pubmed/28525643 http://dx.doi.org/10.1093/nar/gkx298 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Methods Online
Kawasaki, Shunsuke
Fujita, Yoshihiko
Nagaike, Takashi
Tomita, Kozo
Saito, Hirohide
Synthetic mRNA devices that detect endogenous proteins and distinguish mammalian cells
title Synthetic mRNA devices that detect endogenous proteins and distinguish mammalian cells
title_full Synthetic mRNA devices that detect endogenous proteins and distinguish mammalian cells
title_fullStr Synthetic mRNA devices that detect endogenous proteins and distinguish mammalian cells
title_full_unstemmed Synthetic mRNA devices that detect endogenous proteins and distinguish mammalian cells
title_short Synthetic mRNA devices that detect endogenous proteins and distinguish mammalian cells
title_sort synthetic mrna devices that detect endogenous proteins and distinguish mammalian cells
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5499560/
https://www.ncbi.nlm.nih.gov/pubmed/28525643
http://dx.doi.org/10.1093/nar/gkx298
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