短发夹RNA文库及其在癌症遗传学中的应用。

shRNA libraries and their use in cancer genetics.

作者信息

Bernards René, Brummelkamp Thijn R, Beijersbergen Roderick L

机构信息

Division of Molecular Carcinogenesis and Centre for Biomedical Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.

出版信息

Nat Methods. 2006 Sep;3(9):701-6. doi: 10.1038/nmeth921.

DOI:10.1038/nmeth921

PMID:16929315

Abstract

RNA interference was originally described as a powerful tool to inhibit gene expression in model organisms. Until recently, loss-of-function genetic screens in mammalian cells were hampered by a lack of suitable tools that can be used in a high-throughput format. Here we discuss the construction of short-hairpin RNA (shRNA) vector libraries, in particular those generated at the Netherlands Cancer Institute (NKI), and their application in mammalian cancer genetics. We describe their virtues and limitations, as well as different options for screening such libraries.

摘要

RNA干扰最初被描述为一种在模式生物中抑制基因表达的强大工具。直到最近，哺乳动物细胞中的功能丧失型遗传筛选一直受到缺乏可用于高通量形式的合适工具的阻碍。在这里，我们讨论短发夹RNA（shRNA）载体文库的构建，特别是荷兰癌症研究所（NKI）构建的文库，以及它们在哺乳动物癌症遗传学中的应用。我们描述了它们的优点和局限性，以及筛选此类文库的不同选择。