RNAi screening in Drosophila cells and in vivo

Mohr, Stephanie E.

doi:10.1016/j.ymeth.2014.02.018

Cited by 35 publications

(30 citation statements)

References 81 publications

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“…Thus, a number of genes that are non-essential but cause morphological defects are missed in our screen. On the other hand, RNAi may not be efficient or cause off targeting effects (Green et al, 2014; Mohr, 2014). Regardless of the methods that are being used, rescue experiments and independent validation are critical to determine that the phenotype one observes is due to loss of the gene of interest when performing a genetic screen.…”

Section: Discussionmentioning

confidence: 99%

A Drosophila Genetic Resource of Mutants to Study Mechanisms Underlying Human Genetic Diseases

Yamamoto

Jaiswal

Charng

et al. 2014

Cell

352

367

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Summary Invertebrate model systems are powerful tools for studying human disease owing to their genetic tractability and ease of screening. We conducted a mosaic genetic screen of lethal mutations on the Drosophila X-chromosome to identify genes required for the development, function, and maintenance of the nervous system. We identified 165 genes, most of whose function has not been studied in vivo. In parallel, we investigated rare variant alleles in 1,929 human exomes from families with unsolved Mendelian disease. Genes that are essential in flies and have multiple human homologs were found to be likely to be associated with human diseases. Merging the human datasets with the fly genes allowed us to identify disease-associated mutations in six families and to provide insights into microcephaly associated with brain dysgenesis. This bidirectional synergism between fly genetics and human genomics facilitates the functional annotation of evolutionarily conserved genes involved in human health.

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Section: Discussionmentioning

confidence: 99%

A Drosophila Genetic Resource of Mutants to Study Mechanisms Underlying Human Genetic Diseases

Yamamoto

Jaiswal

Charng

et al. 2014

Cell

352

367

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“…He postulated spatially different distribution of surface lipids (especially of high-melting alkanes) in order to explain why water loss rates do not correlate perfectly with the measured 'bulk' lipid-melting point. Thanks to the arsenal of molecular and genetic tools available, including tissue-specific gene silencing methods [34], D. melanogaster is the perfect model insect to uncover the respective underlying genetic and molecular factors. In contrast to the situation in D. melanogaster adults and T. molitor larvae, the surface of locust first-instar nymphs and D. melanogaster larvae is not regionalized.…”

Section: Discussionmentioning

confidence: 99%

Regionalization of surface lipids in insects

Wang

Zhang

et al. 2016

Proc. R. Soc. B.

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Cuticular hydrocarbons (CHCs) play a critical role in the establishment of the waterproof barrier that prevents dehydration and wetting in insects. While rich data are available on CHC composition in different species, we know little about their distribution and organization. Here, we report on our studies of the surface barrier of the fruit fly Drosophila melanogaster applying a newly developed Eosin Y staining method. The inert Eosin Y penetrates different regions of the adult body at distinct temperatures. By contrast, the larval body takes up the dye rather uniformly and gradually with increasing temperature. Cooling down specimens to 258C after incubation at higher temperatures restores impermeability. Eosin Y penetration is also sensitive to lipid solvents such as chloroform indicating that permeability depends on CHCs. As in D. melanogaster adult flies, Eosin Y penetration is regionalized in Tenebrio molitor larvae, whereas it is not in Locusta migratoria nymphs. Regionalization of the fly surface implies tissue-specific variation of the genetic or biochemical programmes of CHC production and deposition. The Eosin Y-based map of CHC distribution may serve to identify the respective factors that are activated to accommodate ecological needs.

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“…RNA interference (RNAi) has been used by numerous Drosophila researchers for experiments in vivo, and in cultured cell lines, and many reagents are available (reviewed in Mohr 2014). Some studies have incorporated RNAi knockdown of DNA repair genes in vivo (e.g., Marek and Bale 2006), and in cells (e.g., Chiolo et al 2011).…”

Section: Other Assaysmentioning

confidence: 99%

DNA Repair inDrosophila: Mutagens, Models, and Missing Genes

2017

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The numerous processes that damage DNA are counterbalanced by a complex network of repair pathways that, collectively, can mend diverse types of damage. Insights into these pathways have come from studies in many different organisms, including Drosophila melanogaster. Indeed, the first ideas about chromosome and gene repair grew out of Drosophila research on the properties of mutations produced by ionizing radiation and mustard gas. Numerous methods have been developed to take advantage of Drosophila genetic tools to elucidate repair processes in whole animals, organs, tissues, and cells. These studies have led to the discovery of key DNA repair pathways, including synthesis-dependent strand annealing, and DNA polymerase theta-mediated end joining. Drosophila appear to utilize other major repair pathways as well, such as base excision repair, nucleotide excision repair, mismatch repair, and interstrand crosslink repair. In a surprising number of cases, however, DNA repair genes whose products play important roles in these pathways in other organisms are missing from the Drosophila genome, raising interesting questions for continued investigations.

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RNAi screening in Drosophila cells and in vivo

Cited by 35 publications

References 81 publications

A Drosophila Genetic Resource of Mutants to Study Mechanisms Underlying Human Genetic Diseases

A Drosophila Genetic Resource of Mutants to Study Mechanisms Underlying Human Genetic Diseases

Regionalization of surface lipids in insects

DNA Repair inDrosophila: Mutagens, Models, and Missing Genes

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