BIOENG-512. Lab methods:
Introduction to the key principles and concepts underlying the screening activity to identify and to characterize bioactive compounds (chemicals, compounds, siRNAs and natural products) acting on a given biological target or a signaling pathway. Applications for early drug discovery, chemical biology and systems biology.
Objectives of the workshop & experimental design
The main goal of this intensive hands-on 1-week course is to familiarize the students with the process of early drug discovery and to practice some of the multidisciplinary techniques associated. We designed experiments to teach and specifically train about the critical steps of assay development, validation for screening and automation with targets selected for allowing a fast comprehension and compatible with the generation of results during the period of the workshop.
COX-2 is the selected target for two assays to be performed by the students, one in vitro and another one cell-based, using the well-known drugs as controls, Non Steroid Anti Inflammatory Drugs, NSAIDs (diclonefac, paracetamol, ibuprofen, etc.). In addition, the cellular system is used for gene-knock down experiments targeting COX-2 and a series of control genes using siRNAs and lipofection as the transfection method. A series of cell viability assays are also included in the program using chemicals and siRNAs probes. A colorimetric end-point method and a fluorescence microscopy approach are utilized as read-outs for these toxicity assays.
Presentation of the drug discovery principles and processes
- Description of the variety of molecular screening assays, from in vitro target-based to cellular phenotypic assays
- Detailed description of the screening activity, high throughput and high content, with special emphasis on the assay development and validation steps
- Generation of experimental results linked to selected assays: in vitro enzymatic, cytotoxicity siRNA cell-transfection assay using end-point and image-based read-outs (HCS).
- Statistical validation of the assays and hits assignment. Data analyses, evaluations and discussion of the generated data in the frame of the screening activity