HIGH-THROUGHPUT DISCOVERY OF INDUCED AND NATURAL POLYMORPHISMS

¹ Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA

² Department of Biology, University of Washington, Seattle, Washington 98195, USA

TILLING (Targeting Induced Local Lesions in Genomes) is a general reverse-genetic strategy that provides an allelic series of induced point mutations in genes of interest. High-throughput TILLING allows the rapid and low-cost discovery of induced point mutations in populations of chemically mutagenized individuals. TILLING can be applied to most species; at our facility in Seattle we are screening a wide range of organisms including Arabidopsis, maize, rice, and zebrafish. While reverse genetic strategies are a powerful tool in understanding gene function, much can be learned from studying natural genetic variation in populations. However, for most species genome sequence was extracted from one or few individuals. Thus there is need for cost-effective tools that can sample variation accurately on a large scale. We have adapted TILLING and developed a method for discovering multiple types of polymorphisms in natural populations, a strategy that we call Ecotilling. As with TILLING, Ecotilling is high-throughput, low-cost and can be applied to most organisms.