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.
Bibliographical noteFunding Information:
This material is based upon work supported by the US Department of Energy, Office of Science, Biological and Environmental Research Program under award DE-SC-0023142. Additionally, M.Z., J.A.C.M. and M.J.S. are supported by the USDA [2018-33522-28747 and 2021-33522-35340] and Advanced Plant Technologies program, DARPA Award [HR001118C0146]; M.Z. is supported by an NIH NIGMS Biotechnology Training grant [NIHT32GM008347]. Funding for open access charge: Discretionary funds.
© 2023 The Author(s).
PubMed: MeSH publication types
- Journal Article
- Research Support, N.I.H., Extramural
- Research Support, U.S. Gov't, Non-P.H.S.
- Research Support, Non-U.S. Gov't