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在哺乳动物中进行基于RNA干扰的大规模选的资源
Patrick J Paddison1, Jose M Silva, Douglas S Conklin
1Cold Spring Harbor Laboratory, Watson School of Biological Sciences, 1 Bungtown Road, Cold Spring Harbor, New York 11724, USA.
Nature
|March 26, 2004
概括
研究人员开发了一个大型RNA干扰 (RNAi) 库,使用短毛针RNAs (shRNAs) 来使哺乳动物的基因沉默. 这种工具有助于基因发现和分析,正如蛋白质酶功能屏幕所示.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 生物信息学是一种生物信息学.
背景情况:
- RNA干扰 (RNAi) 是哺乳动物细胞中基因沉默的一个关键方法.
- 小干扰RNAs (siRNAs) 和短发针RNAs (shRNAs) 广泛用于RNAi.
- 在哺乳动物中进行大规模遗传分析需要有效的工具.
研究的目的:
- 构建和验证用于哺乳动物基因沉默的全面的shRNA表达库.
- 为了实现高通量基因选和基因发现.
- 为功能基因组学研究提供一个有价值的资源.
主要方法:
- 构建一个shRNA表达库,针对超过15,000个人类和小鼠基因.
- 利用复病毒包装,DNA条形码和细菌交配的多功能载体.
- 基因选的应用,以识别人类蛋白质组功能中的缺陷.
主要成果:
- 创建一个库,大约有28000个测序验证的shRNA表达录音带.
- 在功能性遗传屏幕中展示图书馆的实用性.
- 成功识别了参与蛋白质体功能的基因.
结论:
- 开发的大规模RNAi库对于哺乳动物的特定遗传应用是有效的.
- 这个资源有助于基因分析和发现.
- 该shRNA库代表了功能基因组学研究的重大进步.
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