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The explosion of DNA sequence information has created a unique opportunity to investigate the function of genes. The approach that determines the function of genes first defined by DNA sequence analysis, is called reverse genetics. The tools available for reverse genetics of plants include the use of transposons or T-DNA for gene tagging and the use of RNA interference. While powerful, these methods have limitations. For example, they do not work in all plant species.

We have collaborated with the Henikoff lab to develop a method, called Targeting Induced Local Lesions IN Genomes (TILLING), which is reliable and widely applicable. TILLING combines chemical mutagenesis with mutation screens of pooled PCR products, resulting in the isolation of missense and nonsense mutant alleles of the targeted genes.The TILLING technology to discovery and survey of natural variation. The method is called Ecotilling and has been applied to Arabidopsis and human (see reference below).

TILLING has two significant advantages over existing plant gene knock-out tools: first, it is applicable to any plant since it does not require transgenic or cell culture manipulations. Second, it produces an allelic series of mutations including hypomorphic alleles that are useful for genetic analysis. The Seattle TILLING Project (STP) is a FHCRC-UW collaboration that provides Arabidopsis and fruit flies mutants and was operated through a collaboration of the Henikoff and Comai laboratory. With the departure of Luca Comai from Seattle, STP is now operated by Steve Henikoff and Brad Till. The TILLING objectives have been split: Arabidopsis and fruit flies remain at STP. Public service TILLING for rice and wheat is planned at the UC Davis Genome Center. Rice TILLING at UCD is carried our in collaboration with STP, IRRI (Hei Leung), and USDA-ARS at Davis (Thomas Tai). Rice TILLING is funded by USDA-NRI and the Rockefeller Foundation. Wheat TILLING at UCD is carried out in collaboration with the UCD department of Plant Sciences (Jorge Dubcovsky) and it is funded by a USDA-NRI award.

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