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Methods. 2015 Mar;75:96-104. doi: 10.1016/j.ymeth.2015.02.002. Epub 2015 Feb 14.

High-throughput screening approaches to identify regulators of mammalian autophagy.

Author information

1
London Research Institute, Cancer Research UK, Secretory Pathway Laboratory, 44 Lincolns Inn Fields, London, WC2A 3LY, UK.
2
London Research Institute, Cancer Research UK, High-throughput Screening Unit, 44 Lincolns Inn Fields, London, WC2A 3LY, UK.
3
Department of Neurology, The Icahn School of Medicine at Mount Sinai, Hess Center for Science and Medicine, 1470 Madison Avenue, New York, NY 10029, USA.
4
London Research Institute, Cancer Research UK, Secretory Pathway Laboratory, 44 Lincolns Inn Fields, London, WC2A 3LY, UK. Electronic address: Sharon.tooze@cancer.org.uk.

Abstract

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.

KEYWORDS:

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

PMID:
25688674
DOI:
10.1016/j.ymeth.2015.02.002
[Indexed for MEDLINE]

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