Fraser A G, Kamath R S, Zipperlen P, Martinez-Campos M, Sohrmann M, Ahringer J
Wellcome/CRC Institute, University of Cambridge, UK.
Nature. 2000 Nov 16;408(6810):325-30. doi: 10.1038/35042517.
Complete genomic sequence is known for two multicellular eukaryotes, the nematode Caenorhabditis elegans and the fruit fly Drosophila melanogaster, and it will soon be known for humans. However, biological function has been assigned to only a small proportion of the predicted genes in any animal. Here we have used RNA-mediated interference (RNAi) to target nearly 90% of predicted genes on C. elegans chromosome I by feeding worms with bacteria that express double-stranded RNA. We have assigned function to 13.9% of the genes analysed, increasing the number of sequenced genes with known phenotypes on chromosome I from 70 to 378. Although most genes with sterile or embryonic lethal RNAi phenotypes are involved in basal cell metabolism, many genes giving post-embryonic phenotypes have conserved sequences but unknown function. In addition, conserved genes are significantly more likely to have an RNAi phenotype than are genes with no conservation. We have constructed a reusable library of bacterial clones that will permit unlimited RNAi screens in the future; this should help develop a more complete view of the relationships between the genome, gene function and the environment.
两种多细胞真核生物——线虫秀丽隐杆线虫和果蝇黑腹果蝇的完整基因组序列已为人所知，人类的完整基因组序列也将很快知晓。然而，在任何动物中，只有一小部分预测基因的生物学功能已被确定。在这里，我们通过给线虫喂食表达双链RNA的细菌，利用RNA介导的干扰（RNAi）技术靶向秀丽隐杆线虫I号染色体上近90%的预测基因。我们已确定了所分析基因中13.9%的功能，将I号染色体上具有已知表型的已测序基因数量从70个增加到378个。尽管大多数具有不育或胚胎致死RNAi表型的基因参与基础细胞代谢，但许多导致胚胎后期表型的基因具有保守序列，但其功能未知。此外，与无保守性的基因相比，保守基因具有RNAi表型的可能性显著更高。我们构建了一个可重复使用的细菌克隆文库，这将使未来能够进行无限次的RNAi筛选；这应该有助于更全面地了解基因组、基因功能和环境之间的关系。
