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Programmable RNA N6-methyladenosine editing with CRISPR/dCas13a in plants

ABSTRACT: N6-methyladenonsine (m6A) is the most prevalent modification on mRNA and plays critical roles in mRNA processing and metabolism. However, how the m6A modification on individual gene of interest regulates gene function and the links to phenotypic outcome in plants are mostly unknown. Here, we described the construction and characterization of programmable m6A editing tools by fusing the m6A writer, core catalytical domain of MTA and MTB complex, and eraser, ALKBH5, to catalytically dead Cas13a (dCas13a), respectively, for targeting methylation and demethylation of specific mRNA. We demonstrated that our m6A editors could efficiently and specifically add and remove the m6A modification on specific RNA transcript in both Nicotiana benthamiana and Arabidopsis. Moreover, targeting SHORT-ROOT (SHR) transcript with methylation editor could significantly increase its m6A levels with limited off-target effects and enhance its expression, giving rise to induced plant growth with enlarged leaf size and root, increased plant height, plant biomass and total grain weight in Arabidopsis. Collectively, these findings suggest that our programmable m6A editing tools can be applied to study m6A modification of specific genes in plants, and might also have great potential applications for crop improvement in future

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE221143 | GEO | 2024/01/01

REPOSITORIES: GEO

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