MS2 RNA labelling technology for real-time single-cell transcriptional analysis of phosphate and phosphite signalling in plants
Advanced imaging platform combining MS2 RNA labelling, microfluidic devices, and spinning disk microscopy to monitor rapid transcriptional repression in plant cells. Enables precise spatio-temporal characterization of phosphate versus phosphite signalling pathways mediated by SPX inhibitor proteins, supporting antifungal target discovery.
Overview
This solution offers a high-resolution imaging and molecular toolkit for studying rapid transcriptional responses in plant cells, specifically targeting the signalling pathways triggered by phosphate (a macronutrient) and phosphite (an antifungal agent). While both ions repress similar genetic targets, phosphite-induced repression is only transient, and the underlying mechanism remains poorly understood. The platform addresses this gap by enabling real-time, single-cell visualization of transcriptional dynamics that conventional gene reporters (GFP, luciferase) cannot capture due to their slow response kinetics.
The technology supports agricultural biotechnology and crop science applications by providing mechanistic insight into nutrient signalling and antifungal compound action. Understanding the SPX inhibitor family (SPX1–4) and their sequential regulatory activity can inform new antifungal targets and improve fertilizer efficiency strategies.
Technical specifications
Core technology components:
- MS2 RNA labelling system using a fusion between GFP and the MS2 coat protein (MCP-GFP) that recognizes 128 MS2 RNA stem-loops driven by phosphate-repressed promoters
- Microfluidic device integration for precise delivery of phosphate and phosphite treatments at the single-cell level
- Spinning disk confocal microscopy for high-speed, live-cell transcriptional imaging
- Antibody-based detection of SPX inhibitor proteins (SPX1–4) with confirmed post-transcriptional regulation of the main inhibitor
- Recombinant protein expression in bacterial systems for ongoing antibody production
- Western blot analysis workflows to determine repressor protein time windows
Key capabilities:
- Real-time tracking of transcriptional repression spreading through plant tissue within minutes
- Spatial-temporal mapping distinguishing phosphate versus phosphite response kinetics
- Identification of phosphite-specific targets for antifungal development
Technology readiness level
The platform has been validated through proof-of-concept experiments demonstrating transcriptional repression triggered by phosphate addition using the MCP-GFP transgenic line combined with microfluidic and spinning disk imaging. Antibodies for the primary SPX inhibitor have been produced and validated, revealing strong post-transcriptional regulation. Production of additional antibodies for SPX2–4 is underway, with successful bacterial protein expression already achieved. The full comparative study between phosphate and phosphite kinetics is projected to be completed within one year, positioning the technology at an advanced validation stage suitable for collaborative research partnerships.
About Commissariat à l'Énergie Atomique et aux Énergies Alternatives
CEA is a French public research organization with multi‑site campuses and large‑scale scientific infrastructure, operating across the research‑to‑industry continuum with a strong applied focus. Industry engages through co‑located technology institutes, joint laboratories, contract R&D, and access to pilot lines and test facilities on campuses embedded in the Paris‑Saclay and Grenoble innovation clusters. This model supports rapid prototyping, scale‑up, and validation under industrially relevant conditions. Research is funded by the French government alongside competitive awards from national programs and the European Commission. A tech transfer office manages IP, licensing, and venture creation with startup support on major sites.
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