WP1: Identification and function of mobile RNAs delivered to specific cell types and via the phloem

WP1 focuses on the identification of mobile RNAs in the phloem and in single cells of grafted plants, their structural motif(s) triggering transport, and their function as signal under phosphate and sulphate nutrient stresses. We will develop predictive models, using novel single cell transcriptomics pipelines to establish cell-type specific RNA transport and motifs. WP1 includes the following activities:

(a) Identification of phloem miRNA and mRNA populations

(b) Identification of mobile mRNAs in individual cells

(c) Identification and characterisation of motifs triggering RNA mobility

(d) Modelling of inter-organ communication and information flow

WP2: Global identification and characterisation of RNA-binding phloem proteins

WP2 focuses on the identification and characterisation of RBPs that interact specifically with phloem-mobile mRNAs and miRNAs. We aim to characterise their binding affinities, specificity, and their structure to gain insights in the RNA delivery mechanism. WP2 includes the following activities:

(a) Determining the RNA transport facilitators

(b) Structural elucidation of RNP formation

Within both WPs we will adopt novel approaches and techniques that will allow us to build single cell transcriptome profiles and establish the presence of RNAs originating from distant tissues. By using the DESY/ EMBL synchrotron structure elucidation platform, we will be able to resolve the structure of RBPs binding to mobile RNAs and the respective RNP complexes. In both WPs bioinformatics analysis of expression profiles, signalling modelling, and protein and RNA structures are an indispensable part to guarantee success.