We propose to generate, validate and distribute a Xenopus ORFeome, which we define as a fully sequenced, validated set of Xenopus cDNA clones containing one each of every open reading frame (ORF) encoded in the genome, in a format in which any ORF sequence can be easily transferred using recombineering into a diverse array of expression vectors. This one set of reagents will greatly decrease the time to characterize any protein in the myriad of functional assays carried out by the entire Xenopus community. But most importantly, an ORFeome set will allow high-throughput in vivo functional-genomic screening in manner currently not feasible, and will take advantage of a particular strength of the Xenopus system. These clones will be made available, without restriction, to researchers worldwide. The Xenopus community is fully supportive of this project and at a recent PI meeting (Lake Louise, September 2010) the ORFeome was voted as the Top priority of needed Xenopus resources.
We will develop a genomic tool that will greatly enhance the scientific output of the entire scientific community that utilizes the frog Xenopus as a model organism, known as the ORFeome. This will greatly enhance the return of the investment of NIH dollars into Xenopus research which currently contains over 100 laboratories and represents ~$124,000,000 of NIH funding per year. An ORFeome is a highly validated tool that has successfully increased the productivity of human, worm and yeast laboratories. The Xenopus community is fully supportive of this project and at a recent PI meeting (Lake Louise, September 2010) the ORFeome was voted as the Top priority of needed Xenopus resources. Moreover the production of the ORFeome will greatly enhance the usefulness of the current genome information, which is the second highest priority of the community.
|Eagleson, Gerald; Pfister, Katherine; Knowlton, Anne L et al. (2015) Kif2a depletion generates chromosome segregation and pole coalescence defects in animal caps and inhibits gastrulation of the Xenopus embryo. Mol Biol Cell 26:924-37|
|Grant, Ian M; Balcha, Dawit; Hao, Tong et al. (2015) The Xenopus ORFeome: A resource that enables functional genomics. Dev Biol 408:345-57|