High-throughput screening approaches to identify regulators of mammalian autophagy

被引：10

作者：

Joachim, Justin ^{[1
]}

Jiang, Ming ^{[2
]}

McKnight, Nicole C. ^{[3
]}

Howell, Michael ^{[2
]}

Tooze, Sharon A. ^{[1
]}

机构：

[1] Canc Res UK, London Res Inst, Secretory Pathway Lab, London WC2A 3LY, England

[2] Canc Res UK, London Res Inst, High Throughput Screening Unit, London WC2A 3LY, England

[3] Icahn Sch Med Mt Sinai, Dept Neurol, Hess Ctr Sci & Med, New York, NY 10029 USA

来源：

METHODS | 2015年 / 75卷

关键词：

Autophagy; Autophagosome; High-throughput screens; siRNA; Drug and compound libraries; Amino acid starvation; GFP-LC3; p62; Z-score; REVEALS; HOMOLOG; DISEASE; ASSAYS;

D O I：

10.1016/j.ymeth.2015.02.002

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

This article discusses the issues to consider in the development and implementation of high-throughput screens (HTSs) using both siRNA libraries and small molecule compound collections, in order to discover autophagy regulators in mammalian cells. We discuss how to develop the screen, focusing on the key parameters to establish in order to perform a successful screen. As our understanding of autophagy increases and its impact on human disease is elucidated, this technology can be further exploited to uncover novel genes, which may one day become new therapeutic targets. (C) 2015 Elsevier Inc. All rights reserved.

引用

页码：96 / 104

页数：9

共 29 条

[1] Getting ready for building: signaling and autophagosome biogenesis
Abada, Adi
Elazar, Zvulun
[J]. EMBO REPORTS, 2014, 15 (08) : 839 - 852
[2] Screen for Chemical Modulators of Autophagy Reveals Novel Therapeutic Inhibitors of mTORC1 Signaling
Balgi, Aruna D.
Fonseca, Bruno D.
Donohue, Elizabeth
Tsang, Trevor C. F.
Lajoie, Patrick
Proud, Christopher G.
Nabi, Ivan R.
Roberge, Michel
[J]. PLOS ONE, 2009, 4 (09):
[3] Analysis of cell-based RNAi screens
Boutros, Michael
Bras, Ligia P.
Huber, Wolfgang
[J]. GENOME BIOLOGY, 2006, 7 (07)
[4] Buchser W., 2012, ASSAY DEV GUIDELINES
[5] Choi AMK, 2013, NEW ENGL J MED, V368, P651, DOI [10.1056/NEJMra1205406, 10.1056/NEJMc1303158]
[6] Distinct Autophagosomal-Lysosomal Fusion Mechanism Revealed by Thapsigargin-Induced Autophagy Arrest
Ganley, Ian G.
Wong, Pui-Mun
Gammoh, Noor
Jiang, Xuejun
[J]. MOLECULAR CELL, 2011, 42 (06) : 731 - 743
[7] Glucocorticoids downregulate Fyn and inhibit IP3-mediated calcium signaling to promote autophagy in T lymphocytes
Harr, Michael W.
McColl, Karen S.
Zhong, Fei
Molitoris, Jason K.
Distelhorst, Clark W.
[J]. AUTOPHAGY, 2010, 6 (07) : 912 - 921
[8] Tales from an academic RNAi screening facility; FAQs
Jiang, Ming
Instrell, Rachael
Saunders, Becky
Berven, Haakon
Howell, Michael
[J]. BRIEFINGS IN FUNCTIONAL GENOMICS, 2011, 10 (04) : 227 - 237
[9] Selective autophagy mediated by autophagic adapter proteins
Johansen, Terje
Lamark, Trond
[J]. AUTOPHAGY, 2011, 7 (03) : 279 - 296
[10] LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing
Kabeya, Y
Mizushima, N
Uero, T
Yamamoto, A
Kirisako, T
Noda, T
Kominami, E
Ohsumi, Y
Yoshimori, T
[J]. EMBO JOURNAL, 2000, 19 (21) : 5720 - 5728

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