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

New, Maria; Van Acker, Tim; Jiang, Ming; Saunders, Rebecca; Long, Jaclyn S; Sakamaki, Jun-Ichi; Ryan, Kevin M; Howell, Michael; Tooze, Sharon A

New, Maria; Van Acker, Tim; Jiang, Ming; Saunders, Rebecca; Long, Jaclyn S; Sakamaki, Jun-Ichi; Ryan, Kevin M; Howell, Michael; Tooze, Sharon A.

Afiliação

New M; Molecular Cell Biology of Autophagy Laboratory, The Francis Crick Institute, London, UK.
Van Acker T; Molecular Cell Biology of Autophagy Laboratory, The Francis Crick Institute, London, UK.
Jiang M; Tumour Cell Death Laboratory, Cancer Research UK Beatson Institute, Glasgow, UK.
Saunders R; Tumour Cell Death Laboratory, Cancer Research UK Beatson Institute, Glasgow, UK.
Long JS; High Throughput Screening, The Francis Crick Institute, London, UK.
Sakamaki JI; High Throughput Screening, The Francis Crick Institute, London, UK.
Ryan KM; Tumour Cell Death Laboratory, Cancer Research UK Beatson Institute, Glasgow, UK.
Howell M; High Throughput Screening, The Francis Crick Institute, London, UK.
Tooze SA; Tumour Cell Death Laboratory, Cancer Research UK Beatson Institute, Glasgow, UK.

Methods Mol Biol ; 1880: 359-374, 2019.

Article em En | MEDLINE | ID: mdl-30610710

ABSTRACT

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.

Assuntos

Autofagia/genética; Técnicas de Silenciamento de Genes/métodos; RNA Interferente Pequeno/metabolismo; Animais; Técnicas de Cultura de Células/instrumentação; Técnicas de Cultura de Células/métodos; Linhagem Celular; Técnicas de Silenciamento de Genes/instrumentação; Ensaios de Triagem em Larga Escala/instrumentação; Ensaios de Triagem em Larga Escala/métodos; Humanos; Proteínas Associadas aos Microtúbulos/genética; Interferência de RNA; RNA Interferente Pequeno/genética; Transfecção/instrumentação; Transfecção/métodos

Palavras-chave

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

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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Autofagia / RNA Interferente Pequeno / Técnicas de Silenciamento de Genes Tipo de estudo: Diagnostic_studies Limite: Animals / Humans Idioma: En Revista: Methods Mol Biol Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Reino Unido

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