The generation of cellular systems for complex high-throughput screens face very different hurdles, far downstream of substrate generation. The starting cell line often is well established and suitable for automated screening procedures; it is robust enough for reproducible handling independent of the operator, and grows fast enough to be cloned for genetic homogeneity and to provide assay material in reasonable time (for a review, see Chapter 5).
The gene of interest (a receptor, metabolic enzyme or virus protein) can easily be inserted using conventional vectors, transfection methods, and selection systems. Reasonable screening effort yields a cell line of appropriate characteristics with respect to expression level and stability. Challenges arise, however, when downstream effectors, cofactors and detection systems for automated readout have to be stably coexpressed at adjusted levels. Often, analogous and comparable systems must be generated for multiple members of the same protein family, or isoforms of the same protein. Complexity further increases if the screen is directed against interactions among cells or between host cells and parasites in the search for therapeutics against infectious diseases.
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