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.
MeSH terms
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Animals
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Autophagy / genetics*
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Cell Culture Techniques / instrumentation
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Cell Culture Techniques / methods
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Cell Line
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Gene Knockdown Techniques / instrumentation
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Gene Knockdown Techniques / methods*
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High-Throughput Screening Assays / instrumentation
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High-Throughput Screening Assays / methods
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Humans
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Microtubule-Associated Proteins / genetics
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RNA Interference
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism*
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Transfection / instrumentation
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Transfection / methods
Substances
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Microtubule-Associated Proteins
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RNA, Small Interfering