Advanced Graduate Courses: CB 880 Molecular Screening

With the goal of providing hands-on planning, conceptualization, and assay development of a molecular screening strategy relevant to cancer biology and cancer therapeutics, a wet-lab experience will be implemented. This wet-lab component of the course will be centered on the Molecular Screening Facility at The Wistar Institute and will be led by Dr. David Schultz. Dr. Paul Lieberman will supervise this course.

The Molecular Screening Facility maintains libraries and equipment for high-throughput screening of small molecule and nucleic acid (e.g., cDNA, miRNA, siRNA, shRNA) libraries. The course will be based on the optimization and implementation of a real automated assay for high-throughput chemical and functional genomic screens routinely ordered by faculties at Wistar and USciences. The nature of the assay(s) (e.g., biochemical or cell-based) will depend on the requirements of the investigators, but in any case, the experiments may take full advantage of the multiple libraries and equipment available at both institutions.

The process of assay development includes the identification of specific assay types for screening and determination of structure-activity relationships; development of assay reagents; optimization of assay parameters for signal intensity, signal window, and precision; adaptation to automation; scalability; and quantitative assessment of the assay's fitness for screening. Assay parameters include sensitivity to enable identification of compounds with low-potency, precision of biological response between wells and plates, accuracy of positive and negative control compounds with known pharmacology toward the intended target, and economic feasibility.

The goal of this course is to prepare the students for the timely delivery of a robust and well-validated biological assay in screening experiments. Some of the equipment that will be made available to students includes:

- Liquid-handling robotics to transfer compounds/nucleic acids from library stock plates to assay plates

- Automated dispensers to rapidly fill assay plates with bulk reagents (e.g., cells, proteins, buffers)

- Plate reader with absorbance, luminescence, ultraluminescence, fluorescence, fluorescence polarization, time-resolved fluorescence, and alpha screen detection capability

- Automated plate washers

- Tissue culture hood and incubators

- Multichannel pipettes

- Plate sealer and bar code labeler