Poster: Genomics Resources

Abs # 947: High-throughput TILLinG for functional genomics

Presenter:	Greene, Elizabeth A, eagreene@fhcrc.org
Authors	Greene, Elizabeth A (A)   Till, Bradley J (A)   Reynolds, Steven H (B)   Burtner, Chris  (B)   Codomo, Christine A (B)   Enns, Linda C (B)   Henikoff, Jorja G (A)   Johnson, Jessica E (B)   Odden, Anthony R (B)   Taylor, Nicholas E (A)   Young, Kim  (B)   Comai, Luca  (B)   Henikoff, Steven  (A)
Affiliations:	(A): Fred Hutchinson Cancer Research Center (B): Department of Botany, University of Washington
Web Site:	http://tilling.fhcrc.org:9366

TILLinG (Targeting Induced Local Lesions in Genomes) is a general reverse genetic strategy that provides an allelic series of mutations in genes of interest. High-throughput TILLinG allows rapid, low-cost discovery of induced point mutations from a population of chemically mutagenized individuals and is especially well suited to plants where the reference population can be stored as seed. We have developed TILLinG as a service to the Arabidopsis community, as the Arabidopsis TILLinG Project (ATP, http://tilling.fhcrc.org:9366). In its first 18 months of operation ATP has detected, sequenced and delivered ~2000 mutations in 200 genes requested by researchers. Analysis of these data confirms that the mutations available in our reference population are efficiently detected by high-throughput TILLinG, that the full spectrum of point mutations including the more severe deleterious missense and knockout lesions are recovered and that TILLinG permits the recovery of less severe mutations in essential genes. Providing researchers with an allelic series of mutations that range in phenotypic severity permits a more sophisticated analysis of a gene's function. Because high-throughput TILLinG can be applied to any organism which has been mutagenized to a sufficiently high density, we disseminate the technology to the wider genomics community via workshops and the sharing of protocols and informatics. The technology is currently being used in crop plants, in particular in screening mutagenized rice and maize populations. Additionally, in a process called EcoTilling, the high-throughput detection method has been adapted to the discovery of polymorphisms in natural populations, efficiently identifying SNPs, small insertions and deletions and variation in microsatellite repeat number.