Advanced cell biology 1: Experimental approaches

This compulsory course for the "Molecular and Cellular Biology" pathway is designed to deepen and complement the fundamental molecular and cellular processes seen in the BCM2, BCM3 and Advanced Molecular and Cellular Biology courses, by taking a more in-depth look at them. These lessons will focus on the themes of intracellular trafficking, the cell cycle and apoptosis, following on from the BCM3 course.

Students will be asked to analyze a scientific article and produce a structured summary giving the context of the study, the precise scientific question posed by the authors, the strategies implemented and the techniques used to answer it. The aim of this assignment is to help prepare students for writing their TER reports and Master's internships.

TD and TP will be carried out in an integrated way on the same themes as those covered in CM. The practical sessions will be directly linked to the lab work. Students will be asked to pose a question related to a cellular mechanism and, in line with the scientific approach, propose an experimental strategy to answer it. Practical work will enable them to apply this strategy, 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

- Understand the main principles of protein addressing to different cellular compartments.

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

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

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

- Knowledge of approaches for studying modes of interaction between proteins (direct/indirect) (pull down, immunoprecipitation, in vitro interaction, etc.).

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

- 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 summarize them quickly (level 1 article analysis).

- perform plasmid transfection on cell lines.

- know how to perform immunocytochemical/fluorescent labeling.

- know the basics of using an epifluorescence upright microscope.

- use simple image analysis and bioinformatics tools

- People skills :

- work independently and in groups

- Write a report on 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 cell line culture

- From molecules to cells (S1).

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

- Cellular and molecular biology (S3).

- Cellular and molecular biology (S4)

- Advanced cellular and molecular biology (S4)

- This course provides an in-depth understanding of the fundamental concepts and techniques of Cell Biology and Molecular Biology, and is a prerequisite for the Master's degree in Genetics, Epigenetics and Cellular Determinism.