• Login
    Search 
    •   Home
    • Search
    •   Home
    • Search
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Browse

    All of eScholarship@UMassChanCommunitiesPublication DateAuthorsUMass Chan AffiliationsTitlesDocument TypesKeywords

    My Account

    LoginRegister

    Filter by Category

    Date Issued2012 (1)AuthorFreeman, Marc R. (1)Gonzalez, Michael A. (1)
    Hulme, William (1)
    Lebrigio, Rafael F. Acosta (1)Logan, Mary (1)View MoreUMass Chan AffiliationFreeman Lab (1)Graduate School of Biomedical Sciences, Neuroscience Program (1)Neurobiology (1)Document TypeJournal Article (1)KeywordBioinformatics (1)Computational Biology (1)Drosophila melanogaster (1)EMS screen (1)Genetics (1)View MoreJournalBiology (1)

    Help

    AboutSubmission GuidelinesData Deposit PolicySearchingTerms of UseWebsite Migration FAQ

    Statistics

    Most Popular ItemsStatistics by CountryMost Popular Authors
     

    Search

    Show Advanced FiltersHide Advanced Filters

    Filters

    • Publications
    • Profiles

    Now showing items 1-1 of 1

    • List view
    • Grid view
    • Sort Options:
    • Relevance
    • Title Asc
    • Title Desc
    • Issue Date Asc
    • Issue Date Desc
    • Results Per Page:
    • 5
    • 10
    • 20
    • 40
    • 60
    • 80
    • 100

    • 1CSV
    • 1RefMan
    • 1EndNote
    • 1BibTex
    • Selective Export
    • Select All
    • Help
    Thumbnail

    Whole Genome Sequencing and a New Bioinformatics Platform Allow for Rapid Gene Identification in D. melanogaster EMS Screens

    Gonzalez, Michael A.; Van Booven, Derek; Hulme, William; Ulloa, Rick H.; Lebrigio, Rafael F. Acosta; Osterloh, Jeannette M.; Logan, Mary; Freeman, Marc R.; Zuchner, Stephan (2012-12-05)
    Forward genetic screens in Drosophila melanogaster using ethyl methanesulfonate (EMS) mutagenesis are a powerful approach for identifying genes that modulate specific biological processes in an in vivo setting. The mapping of genes that contain randomly-induced point mutations has become more efficient in Drosophila thanks to the maturation and availability of many types of genetic tools. However, classic approaches to gene mapping are relatively slow and ultimately require extensive Sanger sequencing of candidate chromosomal loci. With the advent of new high-throughput sequencing techniques, it is increasingly efficient to directly re-sequence the whole genome of model organisms. This approach, in combination with traditional chromosomal mapping, has the potential to greatly simplify and accelerate mutation identification in mutants generated in EMS screens. Here we show that next-generation sequencing (NGS) is an accurate and efficient tool for high-throughput sequencing and mutation discovery in Drosophila melanogaster. As a test case, mutant strains of Drosophila that exhibited long-term survival of severed peripheral axons were identified in a forward EMS mutagenesis. All mutants were recessive and fell into a single lethal complementation group, which suggested that a single gene was responsible for the protective axon degenerative phenotype. Whole genome sequencing of these genomes identified the underlying gene ect4. To improve the process of genome wide mutation identification, we developed Genomes Management Application (GEM.app, https://genomics.med.miami.edu), a graphical online user interface to a custom query framework. Using a custom GEM.app query, we were able to identify that each mutant carried a unique non-sense mutation in the gene ect4 (dSarm), which was recently shown by Osterloh et al. to be essential for the activation of axonal degeneration. Our results demonstrate the current advantages and limitations of NGS in Drosophila and we introduce GEM.app as a simple yet powerful genomics analysis tool for the Drosophila community. At a current cost of < $1,000 per genome, NGS should thus become a standard gene discovery tool in EMS induced genetic forward screens.
    DSpace software (copyright © 2002 - 2023)  DuraSpace
    Lamar Soutter Library, UMass Chan Medical School | 55 Lake Avenue North | Worcester, MA 01655 USA
    Quick Guide | escholarship@umassmed.edu
    Open Repository is a service operated by 
    Atmire NV
     

    Export search results

    The export option will allow you to export the current search results of the entered query to a file. Different formats are available for download. To export the items, click on the button corresponding with the preferred download format.

    By default, clicking on the export buttons will result in a download of the allowed maximum amount of items.

    To select a subset of the search results, click "Selective Export" button and make a selection of the items you want to export. The amount of items that can be exported at once is similarly restricted as the full export.

    After making a selection, click one of the export format buttons. The amount of items that will be exported is indicated in the bubble next to export format.