Cell microarrays and RNA interference chip away at gene function

Wheeler, Douglas B.; Carpenter, Anne E.; Sabatini, David M.

doi:10.1038/ng1560

Cited by 204 publications

(114 citation statements)

References 65 publications

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“…S1d ) 17,18 . A high-density pattern of siRNA spots, such as 5,000 spots per chip, has been previously demonstrated without any obvious cross-contamination 19 . We addressed the issue of cross-contamination by adjusting the size and number of siRNA spots before large-scale screening.…”

Section: Resultsmentioning

confidence: 99%

Genome-wide functional screening of miR-23b as a pleiotropic modulator suppressing cancer metastasis

et al. 2011

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miRNA globally deregulates human carcinoma. A critical open question is how many miRNAs functionally participate in cancer development, particularly in metastasis. We systematically evaluate the capability of all known human miRNAs to regulate certain metastasis-relevant cell behaviours. To perform the high-throughput screen of miRNAs, which regulate cell migration, we developed a novel self-assembled cell microarray. Here we show that over 20 % of miRNAs have migratory regulation activity in diverse cell types, indicating a general involvement of miRNAs in migratory regulation. MiR-23b, which is downregulated in human colon cancer samples, potently mediates the multiple steps of metastasis, including tumour growth, invasion and angiogenesis in vivo . It regulates a cohort of prometastatic targets, including FZD7 or MAP3k1 . These fi ndings provide new insight into the physiological and potential therapeutic importance of miRNAs as a new class of functional modulators.

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Section: Resultsmentioning

confidence: 99%

Genome-wide functional screening of miR-23b as a pleiotropic modulator suppressing cancer metastasis

et al. 2011

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“…One limitation of monitoring cell cycle progression through DNA content only is the fact that many Drosophila cell lines are polyploid. Finally, a smaller-scale screen has been performed on RNAi cell microarrays where dsRNAs at first spotted at high density on a microscope slide to which cells are subsequently added (Wheeler et al 2005). …”

Section: Rnai Hts Methodsmentioning

confidence: 99%

Applications of High-Throughput RNA Interference Screens to Problems in Cell and Developmental Biology

Perrimon

Mathey-Prévôt

2007

Genetics

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RNA interference (RNAi) in tissue culture cells has emerged as an excellent methodology for identifying gene functions systematically and in an unbiased manner. Here, we describe how RNAi highthroughput screening (HTS) in Drosophila cells are currently being performed and emphasize the strengths and weaknesses of the approach. Further, to demonstrate the versatility of the technology, we provide examples of the various applications of the method to problems in signal transduction and cell and developmental biology. Finally, we discuss emerging technological advances that will extend RNAibased screening methods.

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“…Such complex phenotypes can be detected using for instance cell sorting or high throughput microscopy. Microscopic images can subsequently be analysed by software that can be programmed to extract features such as cell shape, DNA content, subcellular localization of proteins and other parameters (Wheeler et al, 2005;Pepperkok and Ellenberg, 2006;Rines et al, 2006). This combination of single-well screening and reading of complex phenotypes is referred to as 'high content screening' (Neumann et al, 2006).…”

Section: Loss-of-function Genetic Screening Toolsmentioning

confidence: 99%

Loss-of-function genetic screens as a tool to improve the diagnosis and treatment of cancer

Mullenders

Bernards

2009

Oncogene

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A major impediment to the effective treatment of cancer is the molecular heterogeneity of the disease, which is also reflected in an equally diverse pattern of clinical responses to therapy. Currently, only few drugs are available that can be used safely and effectively to treat cancer. To improve this situation, the development of novel and highly specific targets for therapy is of utmost importance. Possibly even more importantly, we need better tools to predict which patients will respond to specific therapies. Such drug response biomarkers will be instrumental to individualize the therapy of patients having seemingly similar cancers. In this study, we discuss how RNA interference-based genetic screens can be used to address these two pressing needs in the care for cancer patients.

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Cell microarrays and RNA interference chip away at gene function

Cited by 204 publications

References 65 publications

Genome-wide functional screening of miR-23b as a pleiotropic modulator suppressing cancer metastasis

Genome-wide functional screening of miR-23b as a pleiotropic modulator suppressing cancer metastasis

Applications of High-Throughput RNA Interference Screens to Problems in Cell and Developmental Biology

Loss-of-function genetic screens as a tool to improve the diagnosis and treatment of cancer

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