Advanced Cell Biology 1: Experimental Approaches

This compulsory course for the "Molecular and Cellular Biology" program is designed to deepen and complete the fundamental molecular and cellular processes seen in the BCM2, BCM3 and "Deepening in Cellular and Molecular Biology" courses, mainly in the form of practical work. These courses will focus on the themes of intracellular trafficking, cell cycle and apoptosis in the continuity of the BCM3 course.

A group work will be done in TD during which the students will have to analyze a scientific article to make a structured summary specifying the context of the study, the precise scientific question asked by the authors, the strategies implemented and the techniques used to answer it. This work is intended to help prepare students for the writing of their TER and internship reports in the Master's program.

The tutorials and practical exercises will be carried out in an integrated manner on the same themes as those dealt with in class. The practical sessions will be directly linked to the laboratory sessions. The students will have to ask a question related to a cellular mechanism and, in the respect of the scientific approach, will propose an experimental strategy allowing to answer it. The practical exercises will allow the application of this strategy by integrating biochemistry, molecular biology and cell biology techniques such as immunoblot, cell culture, immunolabeling and fluorescence microscopy. The results obtained will be analyzed using image analysis techniques and bioinformatics.

- Knowledge:

- knowledge and understanding of the fundamental concepts of cell biology and molecular biology

- To know the main principles of the addressing of proteins to the different cellular compartments.

- Know the main principles of vesicular trafficking (notion of fission and fusion and knowledge of some major proteins involved (Rabs, Arf, SNARE, SNX, BAR proteins...) and nuclear.

- To know the different stages of the cell cycle and the major molecular principles of its regulation (CDK cyclin concepts).

- To know the main pathways leading to cell death (necrosis, apoptosis, anoikis...) and the associated signaling mechanisms.

- Knowledge of approaches to study the modes of interaction between proteins (direct/indirect) (pull down, immunoprecipitation, in vitro interaction ...).

- Know the approaches to evaluate the importance of a protein in a cellular mechanism (siRNA, KO mouse cells, expression of dominant-negative or constitutively active mutants) in relation to the notions of loss or gain of function.

- Knowledge of cell transfection approaches

- Know-how :

- know how to analyze a figure in a scientific article.

- Know how to analyze and interpret the results of a scientific article and be able to make a quick synthesis (level 1 of article analysis).

- know how to perform a plasmid transfection on a cell line.

- know how to perform an imunocytochemical / fluorescent labelling.

- Know the basics of using an epifluorescence upright microscope.

- know how to use simple image analysis and bioinformatics tools

- Know how to be :

- work independently and in groups

- Write a report of practical work respecting the rules of presentation and the time allowed

- respect the hygiene and safety rules of a laboratory and the good laboratory practices related to the culture of cell lines

- From molecules to cells (S1).

- Biochemistry and molecular biology of the cell (S2).

- Cellular and molecular biology (S3).

- Cellular and molecular biology (S4)

- Deepening in cellular and molecular biology (S4)

- This course provides an in-depth study of the fundamental concepts and techniques of cellular and molecular biology and is essential for the master's degree course in Genetics, Epigenetics and Cellular Determinism.