Fluorescence-Based Phenotypic Selection Allows Forward Genetic Screens in Haploid Human Cells
Fluorescence-Based Phenotypic Selection Allows Forward Genetic Screens in Haploid Human Cells
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DOI：
10.1371/journal.pone.0039651
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发表时间：
2012-06-22
期刊：
影响因子：
3.7
通讯作者：
Lehner, Paul J.
中科院分区：
文献类型：
--
作者：
Duncan, Lidia M.;Timms, Richard T.;Lehner, Paul J.
单倍体细胞系的分离最近使得正向遗传筛选的能力能够应用于哺乳动物细胞。如果仅限于致命表型，那么将这种强大的遗传方法应用于哺乳动物系统的兴趣只会因这些筛选的有限效用而减弱。在这里，我们将这些方法的范围扩展到活/死筛选之外，并表明通过荧光激活细胞分选选择细胞表面表型可以识别细胞内途径中的关键分子，在这种情况下是 MHC I 类抗原呈递。非致死单倍体遗传筛选广泛适用于鉴定基本上涉及任何细胞途径的基因。
The isolation of haploid cell lines has recently allowed the power of forward genetic screens to be applied to mammalian cells. The interest in applying this powerful genetic approach to a mammalian system is only tempered by the limited utility of these screens, if confined to lethal phenotypes. Here we expand the scope of these approaches beyond live/dead screens and show that selection for a cell surface phenotype via fluorescence-activated cell sorting can identify the key molecules in an intracellular pathway, in this case MHC class I antigen presentation. Non-lethal haploid genetic screens are widely applicable to identify genes involved in essentially any cellular pathway.