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学科主题基础医学
High-throughput functional screening for autophagy-related genes and identification of TM9SF1 as an autophagosome-inducing gene
He, Pengfei; Peng, Zhi4; Luo, Ye; Wang, Lan2,3; Yu, Peng; Deng, Weiwei; An, Yunqing4; Shi, Taiping1,2,3; Ma, Dalong2,3
关键词high-throughput screening autophagy LC3 TM9SF1 automated fluorescence microscopy system
刊名AUTOPHAGY
2009
5期:1页:52-60
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Cell Biology
研究领域[WOS]Cell Biology
关键词[WOS]MONITORING AUTOPHAGY ; CELL-DEATH ; PROTEIN ; MARKER ; MONODANSYLCADAVERINE ; APOPTOSIS ; HOMOLOG ; MODEL ; YEAST ; LC3
英文摘要

Autophagy, a tightly regulated process responsible for the bulk degradation of most long-lived proteins and some organelles, is associated with several forms of human diseases including cancer, neurodegenerative disease and cardiomyopathies. However, the molecular machinery involved in autophagy in mammalian cells remains poorly understood. Here, we describe a high-throughput, cell-based functional screening platform, based on an automated fluorescence microscopy system, which enables acquiring and quantitatively analyzing images of GFP-LC3 dots in cotransfected cells. From a library of 1,050 human cDNA clones, we identified three genes (TM9SF1, TMEM166 and TMEM74) whose overexpression induced high levels of autophagosome formation. In particular, overexpression of TM9SF1, which colocalized with LC3 according to the confocal assay, led to a significant increase in the number of GFP-LC3 dots. The results of transmission electron microscopy and immunoblotting to examine LC3-II levels further confirmed the ability of TM9SF1 to induce autophagy. Furthermore, knockdown of TM9SF1 expression by RNA interference could hamper starvation-induced autophagy. The functional screening platform therefore can be applied to high-throughput genomic screening candidate autophagy-related genes, which would provide new insights into underlying molecular mechanisms that may regulate autophagy in mammalian cells.

语种英语
WOS记录号WOS:000262844800006
项目编号2006AA02A305
资助机构National High Technology Research and Development Program of China
引用统计
被引频次:43[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
版本出版稿
条目标识符http://ir.bjmu.edu.cn/handle/400002259/65327
专题北京大学基础医学院_免疫学系
作者单位1.Capital Med Univ, Dept Immunol, Beijing, Peoples R China
2.Chinese Natl Human Genome Ctr, Beijing Funct Genom Lab 3, Beijing 100176, Peoples R China
3.Peking Univ, Hlth Sci Ctr, Sch Basic Med Sci, Lab Med Immunol, Beijing 100871, Peoples R China
4.Peking Univ, Ctr Human Dis Genom, Beijing 100871, Peoples R China
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GB/T 7714
He, Pengfei,Peng, Zhi,Luo, Ye,et al. High-throughput functional screening for autophagy-related genes and identification of TM9SF1 as an autophagosome-inducing gene[J]. AUTOPHAGY,2009,5(1):52-60.
APA He, Pengfei.,Peng, Zhi.,Luo, Ye.,Wang, Lan.,Yu, Peng.,...&Ma, Dalong.(2009).High-throughput functional screening for autophagy-related genes and identification of TM9SF1 as an autophagosome-inducing gene.AUTOPHAGY,5(1),52-60.
MLA He, Pengfei,et al."High-throughput functional screening for autophagy-related genes and identification of TM9SF1 as an autophagosome-inducing gene".AUTOPHAGY 5.1(2009):52-60.
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