High-throughput screening approaches to identify regulators of mammalian autophagy

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

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Amino Acids / metabolism
  • Animals
  • Autophagy / drug effects
  • Autophagy / genetics*
  • High-Throughput Screening Assays / methods*
  • Humans
  • Mammals
  • Phagosomes / genetics
  • Phagosomes / metabolism*
  • RNA, Small Interfering / genetics
  • Small Molecule Libraries / pharmacology

Substances

  • Amino Acids
  • RNA, Small Interfering
  • Small Molecule Libraries