INDIVIDUAL RESEARCH PROJECT 8
Imaging cellular heterogeneity in nuclear RNA bursting triggered by Phosphate status variations
Abstract
We want to unravel the variability of the transcription (RNA bursting) between cells of the root during Phosphate homeostasis variation. Combining whole-mount roots growing in a microfluidics system, enabling fine-tuning Pi parameters and MS2/MCP labelling, we will access live, dual imaging (spinning disk microscopy) of the nascent RNAs with a cellular resolution1. 3D single molecule RNA FISH1,2 will be used to validate the results obtained by a distinct approach. Different algorithms (developed by @BNX start-up partners or ChromTrack3D established by WP3) will be used to track the nuclei and identify the transcription site.
The project will have three objectives:
- Quantify the temporal variations of the nascent transcript levels in response to varying Pi status (depletion and refeeding);
- Map the spatial specificity of bursting responses within the different root cell layers
- Quantify the contribution of candidate regulators transcriptional (SPXs repressors of the Pi response) in the bursting process by live-imaging the target transcripts as in Obj1 but in roots deficient for SPXs factors.
More information
Training benefits
Acquisition of a broad experimental background in molecular biology, plant physiology, genetics and cutting edges cell biology approaches:
- Live imaging of the transcription (using MS2/MCP technology) and access to new generation of fluorescent markers
- Dynamic live imaging of protein-protein interactions and signaling molecules (Split FAST technology)
- Microfluidics for live access of molecular modifications taking place in root cells in physiological regulations
Requirements
Good theoretical background in molecular and cell biology. Fluent in English. Experience in plant field would be a bonus.
Environment
The Biosciences and Biotechnologies Institute of Aix-Marseille is located in Provence (FRANCE). It holds state-of-the-art lab facilities allowing full access to the equipment’s required for the project (including top-notch technical Phytotech and Zoom platforms for plant growth (https://www.cite-des-energies.fr/en/home-biam/technological- platforms/phytotec/ or imaging (confocal, spinning disk, spectral and Raman microscopy (https://www.cite- des-energies.fr/en/home-biam/technological-platforms/zoom/).
Responsible PI
ORCID link :
0000-0002-9445-2563
Bibliometry (WoS): 96 articles; >12100 citations; h-index 50. His group raised international recognition in the field of plant Pi homeostasis and devotes part of its efforts to the development of new imaging methodologies to elucidate the complex regulatory mechanisms involved at the cellular level in the control of ion homeostasis.