Longer DNA exhibits greater potential for cell-free gene expression

Abstract Cell-free gene expression systems have been valuable tools for understanding how transcription/translation can be regulated in living cells. Many studies have investigated the determining factors that affect gene expression. Here we report the effect of the length of linearized reporter DNA...

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Autores principales:	Takashi Nishio, Yuko Yoshikawa, Kenichi Yoshikawa, Shin-ichi Sato
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c48755a4b6c0400b946d928e3991b813
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spelling	oai:doaj.org-article:c48755a4b6c0400b946d928e3991b8132021-12-02T17:51:06ZLonger DNA exhibits greater potential for cell-free gene expression10.1038/s41598-021-91243-x2045-2322https://doaj.org/article/c48755a4b6c0400b946d928e3991b8132021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91243-xhttps://doaj.org/toc/2045-2322Abstract Cell-free gene expression systems have been valuable tools for understanding how transcription/translation can be regulated in living cells. Many studies have investigated the determining factors that affect gene expression. Here we report the effect of the length of linearized reporter DNAs encoding the firefly luciferase gene so as to exclude the influence of supercoiling. It is found that longer DNA molecules exhibit significantly greater potency in gene expression; for example, the expression level for DNA with 25.7 kbp is 1000-times higher than that for DNA of 1.7 kbp. AFM observation of the DNA conformation indicates that longer DNA takes shrunken conformation with a higher segment density in the reaction mixture for gene expression, in contrast to the stiff conformation of shorter DNA. We propose an underlying mechanism for the favorable effect of longer DNA on gene expression in terms of the enhancement of access of RNA polymerase to the shrunken conformation. It is expected that the enhancement of gene expression efficiency with a shrunken DNA conformation would also be a rather general mechanism in living cellular environments.Takashi NishioYuko YoshikawaKenichi YoshikawaShin-ichi SatoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-7 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Takashi Nishio Yuko Yoshikawa Kenichi Yoshikawa Shin-ichi Sato Longer DNA exhibits greater potential for cell-free gene expression
description	Abstract Cell-free gene expression systems have been valuable tools for understanding how transcription/translation can be regulated in living cells. Many studies have investigated the determining factors that affect gene expression. Here we report the effect of the length of linearized reporter DNAs encoding the firefly luciferase gene so as to exclude the influence of supercoiling. It is found that longer DNA molecules exhibit significantly greater potency in gene expression; for example, the expression level for DNA with 25.7 kbp is 1000-times higher than that for DNA of 1.7 kbp. AFM observation of the DNA conformation indicates that longer DNA takes shrunken conformation with a higher segment density in the reaction mixture for gene expression, in contrast to the stiff conformation of shorter DNA. We propose an underlying mechanism for the favorable effect of longer DNA on gene expression in terms of the enhancement of access of RNA polymerase to the shrunken conformation. It is expected that the enhancement of gene expression efficiency with a shrunken DNA conformation would also be a rather general mechanism in living cellular environments.
format	article
author	Takashi Nishio Yuko Yoshikawa Kenichi Yoshikawa Shin-ichi Sato
author_facet	Takashi Nishio Yuko Yoshikawa Kenichi Yoshikawa Shin-ichi Sato
author_sort	Takashi Nishio
title	Longer DNA exhibits greater potential for cell-free gene expression
title_short	Longer DNA exhibits greater potential for cell-free gene expression
title_full	Longer DNA exhibits greater potential for cell-free gene expression
title_fullStr	Longer DNA exhibits greater potential for cell-free gene expression
title_full_unstemmed	Longer DNA exhibits greater potential for cell-free gene expression
title_sort	longer dna exhibits greater potential for cell-free gene expression
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/c48755a4b6c0400b946d928e3991b813
work_keys_str_mv	AT takashinishio longerdnaexhibitsgreaterpotentialforcellfreegeneexpression AT yukoyoshikawa longerdnaexhibitsgreaterpotentialforcellfreegeneexpression AT kenichiyoshikawa longerdnaexhibitsgreaterpotentialforcellfreegeneexpression AT shinichisato longerdnaexhibitsgreaterpotentialforcellfreegeneexpression
_version_	1718379311846653952

Longer DNA exhibits greater potential for cell-free gene expression

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