This two-weeks theoretical and practical course focuses on the methods used to study the synthesis, maturation and degradation of a large variety of RNA molecules in eukaryotic cells.
Classically, the diversity of RNA molecules was shared between the well known protein coding messenger RNAs (mRNA) and splicing and translation related tRNA, snRNA, snoRNA or rRNA. The discovery of novel classes of RNA (miRNA, piRNA, CRISPR,…) shed light on new major roles of RNA in cell biology and generated a tremendous amount of interest in RNA related work. Thus, recent developments revealed a crucial role for RNA in regulating gene expression via transcriptional and post-transcriptional control mechanisms. Deep RNA sequencing and high-resolution microarrays also revealed the existence of a highly complex population of non-coding transcripts and sophisticated nuclear and cytoplasmic related surveillance mechanisms. It became thus obvious that organisms have developed several levels of surveillance mechanisms to clean up unwanted transcripts, from aberrant mRNAs to unstable transcripts.
We propose in this practical course to explore in detail the substrates and mechanisms of a major translation-dependent RNA degradation pathway, the Nonsense Mediated mRNA Decay (NMD). NMD degrades transcripts with premature stop codons through poorly understood mechanisms that are conserved from yeast to humans.
The lectures will be delivered by RNA specialists who will focus on the multiple roles of RNA and specific technologies related to RNA studies.
The aim of the practical course is to learn classical and innovative RNA-related techniques that include transcriptome analysis using RNA-Seq, northern blots and RT-qPCR and the study of different RNA populations that physically associate with the NMD machinery. We will compare transcripts from a wild-type strain and from a mutant strain in which an essential NMD factor gene has been deleted. The analysis of genome-wide results will integrate both biochemical association data (co-purification of RNA) and phenotypic data (changes in RNA abundance) to illustrate the diversity of NMD targets.
To get detailed information on the general frame of the course, please refer to the program, that can be downloaded; some topics and practical work schedule change every year.
Candidates must have a good knowledge of genetics, molecular biology and biochemistry (level Master 2), as well as a minimum B1 level in both French and English. Applicants will be evaluated and selected by the course committee.
Unité de Génétique des interactions macromoléculaires
Y. Andéol (Sorbonne Université),
G. Badis-Bréard (Institut Pasteur),
M.-C. Daugeron* (Université Paris Saclay),
M. Fromont-Racine (Institut Pasteur),
A. Jacquier (Institut Pasteur),
M. Lucas-Hourani (Institut Pasteur),
S. Malot (Institut Pasteur),
H. Becker (Sorbonne Université) ,
M. Sala (Institut Pasteur),
H. Waxin (Institut Pasteur),
V. Ponticelli (Institut Pasteur),
C. Torchet (Sorbonne Université),
A. Zider (Université Paris-Diderot).
* Representants des Universités