Efficient gene activation in plants by the MoonTag programmable transcriptional activator

Abstract 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 betw...

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Bibliographic Details
Published in:Nucleic acids research 2023-07, Vol.51 (13), p.7083-7093
Main Authors: Casas-Mollano, J Armando, Zinselmeier, Matthew H, Sychla, Adam, Smanski, Michael J
Format: Article
Language:English
Subjects:
Arabidopsis - genetics
Arabidopsis - metabolism
biotechnology
CRISPR-Cas Systems
CRISPR/Cas9
ENGINEERING
Plant gene activation
Plants, Genetically Modified - genetics
Synthetic Biology and Bioengineering
Transcription Factors - genetics
Transcription Factors - metabolism
Transcriptional Activation
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Summary:Abstract 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. Graphical Abstract Graphical Abstract
ISSN:0305-1048
1362-4962
1362-4962
DOI:10.1093/nar/gkad458