Characterization of Two C. elegans Homologuses of Oncogenic Inhibitor of Apoptosis Proteins (IAPs) and Identification of Interacting Genes.

摘要

Although there is now a phenomenal quantity of sequence data available, biological function has only been assigned to a small percentage of predicted genes in any metazoan. Understanding how genetic information relates to biological function at the level not only of a single gene but of an entire genome is thus a key problem. One approach is to analyse loss-of-function phenotypes of every predicted gene and thus attempt to draw both global and particular conclusions about the relationship between gene sequence, predicted protein product and function in the organism. This is the approach that I have been pursuing for the last 2 years in the nematode worm, C. elegans. We have generated a reagent that uses RNA-mediated interference (RNAi) to individually inhibit -90% of all -19,000 predicted genes in the C. elegans genome simply by feeding dsRNA-expressing bacteria to worms. This technique is both efficient and technically easy, so a single person can analyse loss-of-function phenotypes of up to 1000 genes in a single day for minimal cost; in addition, the reagent can be used an indefinite number of times. In essence, most genetic screens in C. elegans that could hitherto be carried out using standard forward genetics can now be carried out using our RNAi reagent, which has the advantage that any detected phenotype is automatically related to gene sequence, obviating the need for positional cloning.