Silva Jose M, Marran Krista, Parker Joel S, Silva Javier, Golding Michael, Schlabach Michael R, Elledge Stephen J, Hannon Gregory J, Chang Kenneth
Watson School of Biological Sciences, Howard Hughes Medical Institute, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Science. 2008 Feb 1;319(5863):617-20. doi: 10.1126/science.1149185.
By virtue of their accumulated genetic alterations, tumor cells may acquire vulnerabilities that create opportunities for therapeutic intervention. We have devised a massively parallel strategy for screening short hairpin RNA (shRNA) collections for stable loss-of-function phenotypes. We assayed from 6000 to 20,000 shRNAs simultaneously to identify genes important for the proliferation and survival of five cell lines derived from human mammary tissue. Lethal shRNAs common to these cell lines targeted many known cell-cycle regulatory networks. Cell line-specific sensitivities to suppression of protein complexes and biological pathways also emerged, and these could be validated by RNA interference (RNAi) and pharmacologically. These studies establish a practical platform for genome-scale screening of complex phenotypes in mammalian cells and demonstrate that RNAi can be used to expose genotype-specific sensitivities.
由于其累积的基因改变，肿瘤细胞可能会出现一些弱点，从而为治疗干预创造机会。我们设计了一种大规模平行策略，用于筛选短发夹RNA（shRNA）文库，以寻找稳定的功能丧失表型。我们同时检测了6000至20000个shRNA，以鉴定对源自人乳腺组织的五种细胞系的增殖和存活至关重要的基因。这些细胞系共有的致死性shRNA靶向许多已知的细胞周期调控网络。细胞系对蛋白质复合物和生物途径抑制的特异性敏感性也显现出来，并且可以通过RNA干扰（RNAi）和药理学方法进行验证。这些研究建立了一个用于在哺乳动物细胞中进行复杂表型基因组规模筛选的实用平台，并证明RNAi可用于揭示基因型特异性敏感性。
