Gain Of Function Screens Using Mammalian Cdnas

Finally a number of reports have utilized the systematic overexpression of individual genes using arrayed, characterized full length cDNA libraries. These studies identified many novel regulators of a variety of cellular signaling pathways including that regulated by inflammatory cytokines, cAMP, wnt, and AP-1 proteins

[22-24]. While these initial reports used simple reporter gene assays to identify signaling proteins, a recent report [25] utilized a high-throughput microscopy assay to identify proteins capable of inducing the transport of [TORCs] (transducers of regulated CREB) from the cytoplasm to the nucleus. Interestingly, the TORC proteins were themselves previously identified in high-throughput screens using arrayed cDNA libraries. Another study utilized real-time microscopy to identify novel growth regulatory molecules [26]. While overexpression of genes through cDNAs is essentially how all expression cloning was done over the last 20 years, the systematic high-throughput use of this technique appears to be highly effective at identifying novel and specific regulatory proteins in a variety of mammalian systems.

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