It is essential that the Arabidopsis research community finish the job of building its reverse-genetic infrastructure. That infrastructure will not be complete until two important criteria are satisfied: (1) Researchers must have access to a null-mutant of every gene in the genome. (2) Tools must be created that allow tandemly-duplicated genes to be knocked-out. In order to satisfy these needs, a reverse-genetic resource consisting of a collection of mapped T-DNA lines that contain Ds transposon launch-pads and Cre/Lox recombination sites will be developed. These functional elements will give users the ability to create targeted deletions of any segment of the genome, be it a group of tandemly-duplicated genes or a single gene for which no mutant allele currently exists. Through previous efforts, 10,450 of these pDs-Lox T-DNA lines were delivered to the Arabidopsis Biological Resource Center at Ohio State University. The goal of the present project is to deliver an additional 50,000 mapped pDs-Lox T-DNA lines to the Arabidopsis community over the course of the next two years. Details regarding the production and availability of these lines can be found at the following web site: www.biotech.wisc.edu/Arabidopsis. This project will help the Arabidopsis community achieve the goals of the 2010 project by providing scientists with access to null mutants for genes and groups of genes that would otherwise not be available.
Broader Impacts: This community resource will provide scientists with the tools that they need to generate targeted deletions throughout the Arabidopsis genome. In order to ensure that this technology is accessible to the broadest range of researchers, a comprehensive, web-based tutorial will be developed to accompany this new collection of mapped pDs-Lox lines. This tutorial will provide users with both theoretical and practical information relating to the uses of the pDs-Lox population. Detailed lab protocols will be developed that guide users through the following processes: (1) performing genetic crosses; (2) mobilizing a Ds element and screening for re-insertions; (3) performing Cre/Lox recombination and screening for the desired deletion. The availability of these tailor-made protocols will give labs with no prior transposon experience the information that they need to benefit from the pDs-Lox system. This web-based tutorial will be developed by undergraduate students, thereby providing an opportunity for undergraduates to gain valuable experience working as part of a research team.
