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用于永生化细胞和原代细胞基因功能筛选的慢病毒微阵列。

Microarrays of lentiviruses for gene function screens in immortalized and primary cells.

作者信息

Bailey Steve N, Ali Siraj M, Carpenter Anne E, Higgins Caitlin O, Sabatini David M

机构信息

Whitehead Institute for Biomedical Research and Massachusetts Institute of Technology Department of Biology, Nine Cambridge Center, Cambridge, Massachusetts 02142, USA.

出版信息

Nat Methods. 2006 Feb;3(2):117-22. doi: 10.1038/nmeth848.

Abstract

Here we describe lentivirus-infected cell microarrays for the high-throughput screening of gene function in mammalian cells. To create these arrays, we cultured mammalian cells on glass slides 'printed' with lentiviruses pseudotyped as vesicular stomatitis virus glycoprotein, which encode short hairpin RNA or cDNA. Cells that land on the printed 'features' become infected with lentivirus, creating a living array of stably transduced cell clusters within a monolayer of uninfected cells. The small size of the features of the microarrays (300 microm in diameter) allows high-density spotting of lentivirus, permitting thousands of distinct parallel infections on a single glass slide. Because lentiviruses have a wide cellular tropism, including primary cells, lentivirus-infected cell microarrays can be used as a platform for high-throughput screening in a variety of cell types.

摘要

在此,我们描述了用于在哺乳动物细胞中高通量筛选基因功能的慢病毒感染细胞微阵列。为了创建这些阵列,我们在印有伪型为水疱性口炎病毒糖蛋白的慢病毒的玻璃载玻片上培养哺乳动物细胞,这些慢病毒编码短发夹RNA或cDNA。落在印刷“特征”上的细胞会被慢病毒感染,从而在未感染细胞的单层内形成稳定转导细胞簇的活阵列。微阵列特征的小尺寸(直径300微米)允许高密度点样慢病毒,从而在单个玻璃载玻片上实现数千种不同的平行感染。由于慢病毒具有广泛的细胞嗜性,包括原代细胞,慢病毒感染细胞微阵列可作为多种细胞类型高通量筛选的平台。

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