Module 3: RNA imaging and gene expression
Organisers
E. Bertrand, E. Margeat
The aim of the module is to show how imaging technique can help to study gene expression. Imaging techniques proving single molecule resolution in live and fixed cells will be used to measure variability of gene expression across a cell population and across time.
The specific question adressed in the module will be to understand how gene ploidy affects expression noise. The activity of a gene in a human cell line will be followed by three techniques: (i) at the level of the protein product (by RICS, FCS, N&B); (ii) at the mRNA level in fixed cells, by smFISH; (iii) at the level of the nascent RNAs in live cells, by tagging the RNA with MS2-GFP.
Lectures
Lectures of the module will be cover several topic and will by given by:
Practical sessions
E. Bertrand, E. Margeat
The aim of the module is to show how imaging technique can help to study gene expression. Imaging techniques proving single molecule resolution in live and fixed cells will be used to measure variability of gene expression across a cell population and across time.
The specific question adressed in the module will be to understand how gene ploidy affects expression noise. The activity of a gene in a human cell line will be followed by three techniques: (i) at the level of the protein product (by RICS, FCS, N&B); (ii) at the mRNA level in fixed cells, by smFISH; (iii) at the level of the nascent RNAs in live cells, by tagging the RNA with MS2-GFP.
Lectures
Lectures of the module will be cover several topic and will by given by:
- Specific-module lecture 1: E. Bertrand/ E. Margeat
- Specific-module lecture 2: F. Müller (Institut Pasteur, Paris): RNA imaging by smFISH
- Specific-module lecture 3: N. Declerck (CBS, Montpellier): Noise at the protein level
- Specific-module lecture 4: X. Darzacq (UC Berkeley).
Practical sessions
- Session 1= fluctuations of transcriptional activity by live cell RNA imaging
- Session 2= counting of nascent and mature single mRNA by WF and SIM microscopy
- Session 3= protein counting by N&B