Longer DNA Exhibits Greater Potential for Cell-free Gene Expression

Overview

Journal Sci Rep

Specialty Science

Date 2021 Jun 4

PMID 34083658

Citations 7

Authors

Takashi Nishio

Yuko Yoshikawa

Kenichi Yoshikawa

Shin-Ichi Sato

Affiliations

Soon will be listed here.

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.

Citing Articles

Design and Characterization of a Gene-Encoding DNA Nanoparticle in a Cell-Free Transcription-Translation System.

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Characteristic effect of hydroxyurea on the higher-order structure of DNA and gene expression.

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The Anticancer Drug Daunomycin Directly Affects Gene Expression and DNA Structure.

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