ArticlePloS one2011
A novel multiplex cell viability assay for high-throughput RNAi screening.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
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- Integrated-gut-liver-on-a-chip platform as an in vitro human model of non-alcoholic fatty liver disease.Communications biology · 2023Article
- Assaying Proliferation Characteristics of Cells Cultured Under Static Versus Periodic Conditions.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Identification ofInternational journal of molecular sciences · 2022Article
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- Bottom-up assembly of biomedical relevant fully synthetic extracellular vesicles.Science advances · 2021Article
- Multiparametric High-Content Assays to Measure Cell Health and Oxidative Damage as a Model for Drug-Induced Liver Injury.Current protocols in chemical biology · 2020Article
- Synthesis of Tamoxifen-Artemisinin and Estrogen-Artemisinin Hybrids Highly Potent Against Breast and Prostate Cancer.ChemMedChem · 2020Article
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- Proliferation characteristics of cells cultured under periodic versus static conditions.Cytotechnology · 2019Article
- Chemo-photothermal effects of doxorubicin/silica-carbon hollow spheres on liver cancer.RSC advances · 2018Article
- Machine learning and image-based profiling in drug discovery.Current opinion in systems biology · 2018Review
- RNA Interference (RNAi) Screening inGenetics · 2018Article
- Nano- and Micro-Patterned S-, H-, and X-PDMS for Cell-Based Applications: Comparison of Wettability, Roughness, and Cell-Derived Parameters.Frontiers in bioengineering and biotechnology · 2018Article
- A kinome siRNA screen identifies HGS as a potential target for liver cancers with oncogenic mutations in CTNNB1.BMC cancer · 2015Article
- A high-throughput RNAi screen for detection of immune-checkpoint molecules that mediate tumor resistance to cytotoxic T lymphocytes.EMBO molecular medicine · 2015Article
- Viability assays for cells in culture.Journal of visualized experiments : JoVE · 2014Article
- Robust RNAi enhancement via human Argonaute-2 overexpression from plasmids, viral vectors and cell lines.Nucleic acids research · 2013Article
- Luminescent multiplex viability assay for Trypanosoma brucei gambiense.Parasites & vectors · 2013Article
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Authors and funding
9 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cell-based high-throughput RNAi screening has become a powerful research tool in addressing a variety of biological questions. In RNAi screening, one of the most commonly applied assay system is measuring the fitness of cells that is usually quantified using fluorescence, luminescence and absorption-based readouts. These methods, typically implemented and scaled to large-scale screening format, however often only yield limited information on the cell fitness phenotype due to evaluation of a single and indirect physiological indicator. To address this problem, we have established a cell fitness multiplexing assay which combines a biochemical approach and two fluorescence-based assaying methods. We applied this assay in a large-scale RNAi screening experiment with siRNA pools targeting the human kinome in different modified HEK293 cell lines. Subsequent analysis of ranked fitness phenotypes assessed by the different assaying methods revealed average phenotype intersections of 50.7±2.3%-58.7±14.4% when two indicators were combined and 40-48% when a third indicator was taken into account. From these observations we conclude that combination of multiple fitness measures may decrease false-positive rates and increases confidence for hit selection. Our robust experimental and analytical method improves the classical approach in terms of time, data comprehensiveness and cost.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.