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Identification of a Transferrable Terminator Element That Inhibits Small RNA Production and Improves Transgene Expression Levels

The role of terminators is more commonly associated with the polyadenylation and 3′ end formation of new transcripts. Recent evidence, however, suggests that this regulatory region can have a dramatic impact on gene expression. Nonetheless, little is known about the molecular mechanisms leading to t...

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Autores principales: de Felippes, Felipe Fenselau, Shand, Kylie, Waterhouse, Peter M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9149433/
https://www.ncbi.nlm.nih.gov/pubmed/35651775
http://dx.doi.org/10.3389/fpls.2022.877793
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author de Felippes, Felipe Fenselau
Shand, Kylie
Waterhouse, Peter M.
author_facet de Felippes, Felipe Fenselau
Shand, Kylie
Waterhouse, Peter M.
author_sort de Felippes, Felipe Fenselau
collection PubMed
description The role of terminators is more commonly associated with the polyadenylation and 3′ end formation of new transcripts. Recent evidence, however, suggests that this regulatory region can have a dramatic impact on gene expression. Nonetheless, little is known about the molecular mechanisms leading to the improvements associated with terminator usage in plants and the different elements in a plant terminator. Here, we identified an element in the Arabidopsis HSP18.2 terminator (tHSP) to be essential for the high level of expression seen for transgenes under the regulation of this terminator. Our molecular analyses suggest that this newly identified sequence acts to improve transcription termination, leading to fewer read-through events and decreased amounts of small RNAs originating from the transgene. Besides protecting against silencing, the tHSP-derived sequence positively impacts splicing efficiency, helping to promote gene expression. Moreover, we show that this sequence can be used to generate chimeric terminators with enhanced efficiency, resulting in stronger transgene expression and significantly expanding the availability of efficient terminators that can be part of good expression systems. Thus, our data make an important contribution toward a better understanding of plant terminators, with the identification of a new element that has a direct impact on gene expression, and at the same time, creates new possibilities to modulate gene expression via the manipulation of 3′ regulatory regions.
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spelling pubmed-91494332022-05-31 Identification of a Transferrable Terminator Element That Inhibits Small RNA Production and Improves Transgene Expression Levels de Felippes, Felipe Fenselau Shand, Kylie Waterhouse, Peter M. Front Plant Sci Plant Science The role of terminators is more commonly associated with the polyadenylation and 3′ end formation of new transcripts. Recent evidence, however, suggests that this regulatory region can have a dramatic impact on gene expression. Nonetheless, little is known about the molecular mechanisms leading to the improvements associated with terminator usage in plants and the different elements in a plant terminator. Here, we identified an element in the Arabidopsis HSP18.2 terminator (tHSP) to be essential for the high level of expression seen for transgenes under the regulation of this terminator. Our molecular analyses suggest that this newly identified sequence acts to improve transcription termination, leading to fewer read-through events and decreased amounts of small RNAs originating from the transgene. Besides protecting against silencing, the tHSP-derived sequence positively impacts splicing efficiency, helping to promote gene expression. Moreover, we show that this sequence can be used to generate chimeric terminators with enhanced efficiency, resulting in stronger transgene expression and significantly expanding the availability of efficient terminators that can be part of good expression systems. Thus, our data make an important contribution toward a better understanding of plant terminators, with the identification of a new element that has a direct impact on gene expression, and at the same time, creates new possibilities to modulate gene expression via the manipulation of 3′ regulatory regions. Frontiers Media S.A. 2022-05-16 /pmc/articles/PMC9149433/ /pubmed/35651775 http://dx.doi.org/10.3389/fpls.2022.877793 Text en Copyright © 2022 de Felippes, Shand and Waterhouse. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
de Felippes, Felipe Fenselau
Shand, Kylie
Waterhouse, Peter M.
Identification of a Transferrable Terminator Element That Inhibits Small RNA Production and Improves Transgene Expression Levels
title Identification of a Transferrable Terminator Element That Inhibits Small RNA Production and Improves Transgene Expression Levels
title_full Identification of a Transferrable Terminator Element That Inhibits Small RNA Production and Improves Transgene Expression Levels
title_fullStr Identification of a Transferrable Terminator Element That Inhibits Small RNA Production and Improves Transgene Expression Levels
title_full_unstemmed Identification of a Transferrable Terminator Element That Inhibits Small RNA Production and Improves Transgene Expression Levels
title_short Identification of a Transferrable Terminator Element That Inhibits Small RNA Production and Improves Transgene Expression Levels
title_sort identification of a transferrable terminator element that inhibits small rna production and improves transgene expression levels
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9149433/
https://www.ncbi.nlm.nih.gov/pubmed/35651775
http://dx.doi.org/10.3389/fpls.2022.877793
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