Approach for functional analysis of glycan using RNA interference
Approach for functional analysis of glycan using RNA interference
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DOI：
10.1023/b:glyc.0000043750.80389.14
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发表时间：
2004-01-01
影响因子：
3
通讯作者：
Nakamura, M
中科院分区：
文献类型：
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作者：
Nishihara, S;Ueda, R;Nakamura, M
阐明聚糖的生物学作用是继基因组计划之后需要解决的最重要的问题之一。RNA干扰正成为研究模式生物基因功能的一种有效的反向遗传学工具。线虫和黑腹果蝇。我们的分子进化研究表明，一个原型的糖基转移酶，在高尔基体中合成各种聚糖结构，是哺乳动物和果蝇之间的保守。为了分析聚糖的基本生理功能，我们建立了果蝇诱导型RNAi敲除系统，并将其分别应用于蛋白聚糖UDP-半乳糖：β-木糖1，4-半乳糖基转移酶I和PAPS-转运蛋白。如果在细胞质肌动蛋白启动子的控制下，每个基因的沉默被普遍诱导，RNAi敲低的苍蝇死亡，那么蛋白质是生命不可或缺的。目的基因的表达被特异性干扰，干扰程度与表型密切相关。利用GAL 4-UAS系统获得的可诱导RNAi敲除果蝇为聚糖的功能分析铺平了道路。
The elucidation of the biological role of glycan is one of the most important issues to be resolved following the genome project. RNA interference is becoming an efficient reverse genetic tool for studying gene function in model organisms, including C. elegans and Drosophila melanogaster. Our molecular evolutionary study has shown that a prototype of glycosyltransferases, which synthesize a variety of glycan structures in the Golgi apparatus, was conserved between mammals and Drosophila. For analyses of the basic physiological functions of glycans, we established the Drosophila inducible RNAi knockdown system and applied it to one glycosyltransferase and one transporter, proteoglycan UDP-galactose: beta-xylose 1,4galactosyltransferase I and the PAPS-transporter, respectively. If on the silencing of each gene induced ubiquitously under the control of a cytoplasmic actin promoter, the RNAi knockdown fly died, then the protein was indispensable for life. The expression of the target gene was disrupted specifically and the degree of interference was well correlated with the phenotype. The inducible RNAi knockdown fly obtained using the GAL4-UAS system will pave the way for the functional analysis of glycans.