果蝇细胞生长与活力的全基因组RNA干扰分析

Genome-wide RNAi analysis of growth and viability in Drosophila cells.

作者信息

Boutros Michael, Kiger Amy A, Armknecht Susan, Kerr Kim, Hild Marc, Koch Britta, Haas Stefan A, Paro Renato, Perrimon Norbert

机构信息

Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.

出版信息

Science. 2004 Feb 6;303(5659):832-5. doi: 10.1126/science.1091266.

DOI:10.1126/science.1091266

PMID:14764878

Abstract

A crucial aim upon completion of whole genome sequences is the functional analysis of all predicted genes. We have applied a high-throughput RNA-interference (RNAi) screen of 19,470 double-stranded (ds) RNAs in cultured cells to characterize the function of nearly all (91%) predicted Drosophila genes in cell growth and viability. We found 438 dsRNAs that identified essential genes, among which 80% lacked mutant alleles. A quantitative assay of cell number was applied to identify genes of known and uncharacterized functions. In particular, we demonstrate a role for the homolog of a mammalian acute myeloid leukemia gene (AML1) in cell survival. Such a systematic screen for cell phenotypes, such as cell viability, can thus be effective in characterizing functionally related genes on a genome-wide scale.

摘要

完成全基因组测序后的一个关键目标是对所有预测基因进行功能分析。我们在培养细胞中对19470条双链（ds）RNA进行了高通量RNA干扰（RNAi）筛选，以表征几乎所有（91%）预测的果蝇基因在细胞生长和活力方面的功能。我们发现了438条能鉴定出必需基因的dsRNA，其中80%没有突变等位基因。应用细胞数量的定量测定来鉴定已知和未知功能的基因。特别是，我们证明了哺乳动物急性髓系白血病基因（AML1）的同源物在细胞存活中的作用。因此，这样一个针对细胞表型（如细胞活力）的系统筛选，在全基因组范围内表征功能相关基因方面可能是有效的。