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Efficient gene activation in plants by the MoonTag programmable transcriptional activator
CRISPR/Cas-based transcriptional activators have been developed to induce gene expression in eukaryotic and prokaryotic organisms. The main advantages of CRISPR/Cas-based systems is that they can achieve high levels of transcriptional activation and are very easy to program via pairing between the g...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10359618/ https://www.ncbi.nlm.nih.gov/pubmed/37254802 http://dx.doi.org/10.1093/nar/gkad458 |
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author | Casas-Mollano, J Armando Zinselmeier, Matthew H Sychla, Adam Smanski, Michael J |
author_facet | Casas-Mollano, J Armando Zinselmeier, Matthew H Sychla, Adam Smanski, Michael J |
author_sort | Casas-Mollano, J Armando |
collection | PubMed |
description | CRISPR/Cas-based transcriptional activators have been developed to induce gene expression in eukaryotic and prokaryotic organisms. The main advantages of CRISPR/Cas-based systems is that they can achieve high levels of transcriptional activation and are very easy to program via pairing between the guide RNA and the DNA target strand. SunTag is a second-generation system that activates transcription by recruiting multiple copies of an activation domain (AD) to its target promoters. SunTag is a strong activator; however, in some species it is difficult to stably express. To overcome this problem, we designed MoonTag, a new activator that works on the same basic principle as SunTag, but whose components are better tolerated when stably expressed in transgenic plants. We demonstrate that MoonTag is capable of inducing high levels of transcription in all plants tested. In Setaria, MoonTag is capable of inducing high levels of transcription of reporter genes as well as of endogenous genes. More important, MoonTag components are expressed in transgenic plants to high levels without any deleterious effects. MoonTag is also able to efficiently activate genes in eudicotyledonous species such as Arabidopsis and tomato. Finally, we show that MoonTag activation is functional across a range of temperatures, which is promising for potential field applications. |
format | Online Article Text |
id | pubmed-10359618 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-103596182023-07-22 Efficient gene activation in plants by the MoonTag programmable transcriptional activator Casas-Mollano, J Armando Zinselmeier, Matthew H Sychla, Adam Smanski, Michael J Nucleic Acids Res Synthetic Biology and Bioengineering CRISPR/Cas-based transcriptional activators have been developed to induce gene expression in eukaryotic and prokaryotic organisms. The main advantages of CRISPR/Cas-based systems is that they can achieve high levels of transcriptional activation and are very easy to program via pairing between the guide RNA and the DNA target strand. SunTag is a second-generation system that activates transcription by recruiting multiple copies of an activation domain (AD) to its target promoters. SunTag is a strong activator; however, in some species it is difficult to stably express. To overcome this problem, we designed MoonTag, a new activator that works on the same basic principle as SunTag, but whose components are better tolerated when stably expressed in transgenic plants. We demonstrate that MoonTag is capable of inducing high levels of transcription in all plants tested. In Setaria, MoonTag is capable of inducing high levels of transcription of reporter genes as well as of endogenous genes. More important, MoonTag components are expressed in transgenic plants to high levels without any deleterious effects. MoonTag is also able to efficiently activate genes in eudicotyledonous species such as Arabidopsis and tomato. Finally, we show that MoonTag activation is functional across a range of temperatures, which is promising for potential field applications. Oxford University Press 2023-05-31 /pmc/articles/PMC10359618/ /pubmed/37254802 http://dx.doi.org/10.1093/nar/gkad458 Text en © The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://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 | Synthetic Biology and Bioengineering Casas-Mollano, J Armando Zinselmeier, Matthew H Sychla, Adam Smanski, Michael J Efficient gene activation in plants by the MoonTag programmable transcriptional activator |
title | Efficient gene activation in plants by the MoonTag programmable transcriptional activator |
title_full | Efficient gene activation in plants by the MoonTag programmable transcriptional activator |
title_fullStr | Efficient gene activation in plants by the MoonTag programmable transcriptional activator |
title_full_unstemmed | Efficient gene activation in plants by the MoonTag programmable transcriptional activator |
title_short | Efficient gene activation in plants by the MoonTag programmable transcriptional activator |
title_sort | efficient gene activation in plants by the moontag programmable transcriptional activator |
topic | Synthetic Biology and Bioengineering |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10359618/ https://www.ncbi.nlm.nih.gov/pubmed/37254802 http://dx.doi.org/10.1093/nar/gkad458 |
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