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Title:
Simultaneous analysis of large-scale RNAi screens for pathogen entry | BMC Genomics
Description:
Background Large-scale RNAi screening has become an important technology for identifying genes involved in biological processes of interest. However, the quality of large-scale RNAi screening is often deteriorated by off-targets effects. In order to find statistically significant effector genes for pathogen entry, we systematically analyzed entry pathways in human host cells for eight pathogens using image-based kinome-wide siRNA screens with siRNAs from three vendors. We propose a Parallel Mixed Model (PMM) approach that simultaneously analyzes several non-identical screens performed with the same RNAi libraries. Results We show that PMM gains statistical power for hit detection due to parallel screening. PMM allows incorporating siRNA weights that can be assigned according to available information on RNAi quality. Moreover, PMM is able to estimate a sharedness score that can be used to focus follow-up efforts on generic or specific gene regulators. By fitting a PMM model to our data, we found several novel hit genes for most of the pathogens studied. Conclusions Our results show parallel RNAi screening can improve the results of individual screens. This is currently particularly interesting when large-scale parallel datasets are becoming more and more publicly available. Our comprehensive siRNA dataset provides a public, freely available resource for further statistical and biological analyses in the high-content, high-throughput siRNA screening field.
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Keywords {🔍}
pubmed, screens, data, pmm, cells, pathogens, infection, gene, genes, sirna, article, pathogen, hits, google, scholar, sirnas, hit, figure, screening, pbs, analysis, rnai, cas, performed, cell, methods, parallel, set, central, number, ranking, shared, shows, results, staining, statistical, libraries, file, effects, model, min, power, monocytogenes, additional, mtt, single, image, fixed, mixed, false,
Topics {✒️}
high-throughput rnai screening large-scale rnai screening high-content rnai screening genome-scale rnai screening additional high-throughput data rna replication-center assay large-scale rnai screens numerous large-scale screens article download pdf large-scale sirna screens functional tgf-beta receptor large-scale parallel datasets infectx high-content datasets green outlines high-light linear mixed-effects models high-performance computing resources anr grant mie-2009-signrupvac open access license high-content screening serine/threonine protein kinase linear mixed model wolf-dietrich hardt immunolgie de marseille-luminy infectx data-access page ceramide-enriched membrane platforms 5 μl rnase-free ddh2o large-scale inference rnai-transfected cell plates mann–whitney-u-test host factors reveals shigella flexneri-specific protocol kinome-wide pathogen screens ordinary linear model heat map shows vaccinia virus-specific protocol full size image 𝜎 𝜖 2 bmc genomics 15 genome-wide resources salmonella typhimurium-specific protocol discover shared mechanisms raquel conde-alvarez anti-gfp antibody actin-based active part assessing differential expression hela ccl-2 cells regulates cell growth cross-validated sirna sets people cofund program access data aggregates
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headline:Simultaneous analysis of large-scale RNAi screens for pathogen entry
description:Large-scale RNAi screening has become an important technology for identifying genes involved in biological processes of interest. However, the quality of large-scale RNAi screening is often deteriorated by off-targets effects. In order to find statistically significant effector genes for pathogen entry, we systematically analyzed entry pathways in human host cells for eight pathogens using image-based kinome-wide siRNA screens with siRNAs from three vendors. We propose a Parallel Mixed Model (PMM) approach that simultaneously analyzes several non-identical screens performed with the same RNAi libraries. We show that PMM gains statistical power for hit detection due to parallel screening. PMM allows incorporating siRNA weights that can be assigned according to available information on RNAi quality. Moreover, PMM is able to estimate a sharedness score that can be used to focus follow-up efforts on generic or specific gene regulators. By fitting a PMM model to our data, we found several novel hit genes for most of the pathogens studied. Our results show parallel RNAi screening can improve the results of individual screens. This is currently particularly interesting when large-scale parallel datasets are becoming more and more publicly available. Our comprehensive siRNA dataset provides a public, freely available resource for further statistical and biological analyses in the high-content, high-throughput siRNA screening field.
datePublished:2014-12-22T00:00:00Z
dateModified:2014-12-22T00:00:00Z
pageStart:1
pageEnd:18
license:https://creativecommons.org/publicdomain/zero/1.0/
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keywords:
High-throughput high-content RNAi screening
Pathogen entry
Linear mixed model
Hit detection
Life Sciences
general
Microarrays
Proteomics
Animal Genetics and Genomics
Microbial Genetics and Genomics
Plant Genetics and Genomics
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headline:Simultaneous analysis of large-scale RNAi screens for pathogen entry
description:Large-scale RNAi screening has become an important technology for identifying genes involved in biological processes of interest. However, the quality of large-scale RNAi screening is often deteriorated by off-targets effects. In order to find statistically significant effector genes for pathogen entry, we systematically analyzed entry pathways in human host cells for eight pathogens using image-based kinome-wide siRNA screens with siRNAs from three vendors. We propose a Parallel Mixed Model (PMM) approach that simultaneously analyzes several non-identical screens performed with the same RNAi libraries. We show that PMM gains statistical power for hit detection due to parallel screening. PMM allows incorporating siRNA weights that can be assigned according to available information on RNAi quality. Moreover, PMM is able to estimate a sharedness score that can be used to focus follow-up efforts on generic or specific gene regulators. By fitting a PMM model to our data, we found several novel hit genes for most of the pathogens studied. Our results show parallel RNAi screening can improve the results of individual screens. This is currently particularly interesting when large-scale parallel datasets are becoming more and more publicly available. Our comprehensive siRNA dataset provides a public, freely available resource for further statistical and biological analyses in the high-content, high-throughput siRNA screening field.
datePublished:2014-12-22T00:00:00Z
dateModified:2014-12-22T00:00:00Z
pageStart:1
pageEnd:18
license:https://creativecommons.org/publicdomain/zero/1.0/
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keywords:
High-throughput high-content RNAi screening
Pathogen entry
Linear mixed model
Hit detection
Life Sciences
general
Microarrays
Proteomics
Animal Genetics and Genomics
Microbial Genetics and Genomics
Plant Genetics and Genomics
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