Project by Asako Sugimoto's group

Large-scale analysis of gene function in Caenorhabditis elegans by high-throughput RNAi.

Ikuma Maeda1, Yuji Kohara2,3, Masayuki Yamamoto1, and Asako Sugimoto1,4

1Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo 113-0032, JAPAN.
2Genome Biology Lab, National Institute of Genetics, Mishima, Shizuoka, 411-8540, JAPAN.
3CREST, Japan Science and Technology Corporation.
4PRESTO, Japan Science and Technology Corporation.



Genome-wide analysis of gene function is essential for the post-genome era, and development of efficient and economical technology suitable for it has been in demand. Here we report a large-scale inactivation of the expressed genes in the nematode Caenorhabditis elegans. For this purpose, we have established a high-throughput "RNAi by soaking" methodology by modifying the conventional RNAi method. A set of tag-sequenced non-redundant cDNAs corresponding to approximately 10,000 genes (representing half of the predicted genes) was used for the systematic RNAi analysis. We processed approximately 2,500 genes to date. 28% of them showed detectable phenotypes in development, such as embryonic lethality, post-embryonic lethality, sterility, and morphological abnormality. Of these, we analyzed the phenotypes of F1 sterility in detail, and we have identified 24 genes that might play important roles in germline development. Combined with the ongoing analysis of expression patterns of these cDNAs, the functional information obtained in this work will provide a starting point for further analysis of each gene. Another finding from this screening is that, the incident of essential genes is significantly lower in the X chromosome than in the autosomes.

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