Chapter title |
The Ancient Genetic Networks of Obesity: Whole-Animal Automated Screening for Conserved Fat Regulators
|
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Book title |
Phenotypic Screening
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Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7847-2_10 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7846-5, 978-1-4939-7847-2
|
Authors |
Wenfan Ke, Anna Drangowska-Way, Daniel Katz, Karsten Siller, Eyleen J. O’Rourke |
Abstract |
Caenorhabditis elegans is the first and only metazoan model that enables whole-body gene knockdown by simply feeding their standard laboratory diet, E. coli, carrying RNA interference (RNAi)-expressing constructs. The simplicity of the RNAi treatment, small size, and fast reproduction rate of C. elegans allow us to perform whole-animal high-throughput genetic screens in wild-type, mutant, or otherwise genetically modified C. elegans. In addition, more than 65% of C. elegans genes are conserved in mammals including human. In particular, C. elegans metabolic pathways are highly conserved, which supports the study of complex diseases such as obesity in this genetically tractable model system. In this chapter, we present a detailed protocol for automated high-throughput whole-animal RNAi screening to identify the pathways promoting obesity in diet-induced and genetically driven obese C. elegans. We describe an optimized high-content screening protocol to score fat mass and body fat distribution in whole animals at large scale. We provide optimized pipelines to automatically score phenotypes using the open-source CellProfiler platform within the context of supercomputer clusters. Further, we present a guideline to optimize information workflow from the automated microscope to a searchable database. The approaches described here enable unveiling the whole network of gene-gene and gene-environment interactions that define metabolic health or disease status in this proven model of human disease, but similar principles can be applied to other disease models. |
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United States | 2 | 40% |
Unknown | 3 | 60% |
Demographic breakdown
Type | Count | As % |
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Members of the public | 4 | 80% |
Scientists | 1 | 20% |
Mendeley readers
Geographical breakdown
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Unknown | 14 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Researcher | 3 | 21% |
Student > Doctoral Student | 1 | 7% |
Student > Bachelor | 1 | 7% |
Unspecified | 1 | 7% |
Student > Ph. D. Student | 1 | 7% |
Other | 3 | 21% |
Unknown | 4 | 29% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 2 | 14% |
Chemical Engineering | 1 | 7% |
Environmental Science | 1 | 7% |
Unspecified | 1 | 7% |
Agricultural and Biological Sciences | 1 | 7% |
Other | 4 | 29% |
Unknown | 4 | 29% |