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Methods Mol Biol. 2019;1880:359-374. doi: 10.1007/978-1-4939-8873-0_24.

Identification and Validation of Novel Autophagy Regulators Using an Endogenous Readout siGENOME Screen.

Author information

1
Molecular Cell Biology of Autophagy Laboratory, The Francis Crick Institute, London, UK.
2
Tumour Cell Death Laboratory, Cancer Research UK Beatson Institute, Glasgow, UK.
3
High Throughput Screening, The Francis Crick Institute, London, UK.
4
Molecular Cell Biology of Autophagy Laboratory, The Francis Crick Institute, London, UK. Sharon.tooze@crick.ac.uk.

Abstract

Autophagy is a highly regulated process, and its deregulation can contribute to various diseases, including cancer, immune diseases, and neurodegenerative disorders. Here we describe the design, protocol, and analysis of an imaging-based high-throughput screen with an endogenous autophagy readout. The screen uses a genome-wide siRNA library to identify autophagy regulators in mammalian cells.

KEYWORDS:

Autophagy; Autophagy regulation; Endogenous readout; Hit identification; LC3 puncta; Large data set analysis; Screen analysis; siGENOME screen

PMID:
30610710
DOI:
10.1007/978-1-4939-8873-0_24
[Indexed for MEDLINE]

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