A simplified method to produce mRNAs and functional proteins from synthetic double-stranded DNA templates

We present a method to synthesize mRNAs from synthetic DNA templates that produce biologically active proteins. To illustrate utility, we constructed five unique synthetic DNA templates, produced mRNAs and demonstrated biologic activity of their translated proteins. Examples include secreted lucifer...

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Main Authors: John T Tossberg, Tashawna M Esmond, Thomas M Aune
Format: Article
Language:English
Published: Taylor & Francis Group 2020-10-01
Series:BioTechniques
Subjects:
Online Access:https://www.future-science.com/doi/10.2144/btn-2020-0037
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author John T Tossberg
Tashawna M Esmond
Thomas M Aune
author_facet John T Tossberg
Tashawna M Esmond
Thomas M Aune
author_sort John T Tossberg
collection DOAJ
description We present a method to synthesize mRNAs from synthetic DNA templates that produce biologically active proteins. To illustrate utility, we constructed five unique synthetic DNA templates, produced mRNAs and demonstrated biologic activity of their translated proteins. Examples include secreted luciferase, enhanced green fluorescence protein, IL-4, and IL-12A and IL-12B to form active IL-12. We propose that this method offers a cost- and time-saving alternative to plasmid-based cloning.
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1940-9818
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publishDate 2020-10-01
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series BioTechniques
spelling doaj-art-6cbccd87a45f401fa875c41b0b7e5d4a2025-08-20T02:26:02ZengTaylor & Francis GroupBioTechniques0736-62051940-98182020-10-0169428128810.2144/btn-2020-0037A simplified method to produce mRNAs and functional proteins from synthetic double-stranded DNA templatesJohn T Tossberg0Tashawna M Esmond1Thomas M Aune21Department of Medicine & Vanderbilt University Medical Center, Nashville, TN 37232, USA1Department of Medicine & Vanderbilt University Medical Center, Nashville, TN 37232, USA1Department of Medicine & Vanderbilt University Medical Center, Nashville, TN 37232, USAWe present a method to synthesize mRNAs from synthetic DNA templates that produce biologically active proteins. To illustrate utility, we constructed five unique synthetic DNA templates, produced mRNAs and demonstrated biologic activity of their translated proteins. Examples include secreted luciferase, enhanced green fluorescence protein, IL-4, and IL-12A and IL-12B to form active IL-12. We propose that this method offers a cost- and time-saving alternative to plasmid-based cloning.https://www.future-science.com/doi/10.2144/btn-2020-0037alternative to molecular cloningcell transfectionchemical DNA synthesisprotein translationsynthetic mRNA from DNA template
spellingShingle John T Tossberg
Tashawna M Esmond
Thomas M Aune
A simplified method to produce mRNAs and functional proteins from synthetic double-stranded DNA templates
BioTechniques
alternative to molecular cloning
cell transfection
chemical DNA synthesis
protein translation
synthetic mRNA from DNA template
title A simplified method to produce mRNAs and functional proteins from synthetic double-stranded DNA templates
title_full A simplified method to produce mRNAs and functional proteins from synthetic double-stranded DNA templates
title_fullStr A simplified method to produce mRNAs and functional proteins from synthetic double-stranded DNA templates
title_full_unstemmed A simplified method to produce mRNAs and functional proteins from synthetic double-stranded DNA templates
title_short A simplified method to produce mRNAs and functional proteins from synthetic double-stranded DNA templates
title_sort simplified method to produce mrnas and functional proteins from synthetic double stranded dna templates
topic alternative to molecular cloning
cell transfection
chemical DNA synthesis
protein translation
synthetic mRNA from DNA template
url https://www.future-science.com/doi/10.2144/btn-2020-0037
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