Plant & Animal Genomes XI Conference Plant & Animal Genomes XI ConferenceJanuary 11-15, 2003
Town & Country Convention Center
San Diego, CA
Workshop: Arabidopsis
In the present genomics era, powerful reverse genetic strategies are needed for elucidating gene and protein function in the context of a whole organism. However, most current reverse genetics strategies lack the generality and high-throughput potential of descriptive genomic approaches, such as methods that rely on microarray hybridization. For example, in plant research, effective insertional mutagenesis and transgenic methods are limited to relatively few species or are inefficient. Fortunately, single-nucleotide changes can be induced in practically any organism using traditional chemical mutagens that have been widely used by geneticists for several decades, and progress has been made in efficiently detecting changes. We have introduced a reverse genetics strategy we call TILLING (for Targeting Induced Local Lesions IN Genomes) based on deleterious point mutations. TILLING uses chemical mutagenesis of reference individuals and screening for point mutations in a region of interest. Because base substitutions in proteins provide allelic series, and not just knockouts, this strategy can yield refined insights into protein function. We have developed a high-throughput TILLING strategy, and have demonstrated its efficacy by establishing the Arabidopsis TILLING Project (http://tilling.fhcrc.org:9366), which provides point mutations as a service for the general Arabidopsis community. We have also begun to disseminate the technology to the wider genomics community via workshops and sharing of protocols. The generality of the TILLING strategy means that it can be usefully applied in agriculture and medical research, and TILLING projects are currently under way for crop plants and model organisms.