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Methods. 2014 Jun 15;68(1):15-28. doi: 10.1016/j.ymeth.2014.02.025. Epub 2014 Feb 28.

Chemical mutagens, transposons, and transgenes to interrogate gene function in Drosophila melanogaster.

Author information

  • 1Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Program in Developmental Biology, Baylor College of Medicine, TX 77030, United States. Electronic address: kv134369@bcm.edu.
  • 2Program in Developmental Biology, Departments of Molecular and Human Genetics, Department of Neuroscience, Howard Hughes Medical Institute, Jan and Dan Duncan Neurological Research Institute, Baylor College of Medicine, Houston, TX 77030, United States. Electronic address: hbellen@bcm.edu.

Abstract

The study of genetics, genes, and chromosomal inheritance was initiated by Thomas Morgan in 1910, when the first visible mutations were identified in fruit flies. The field expanded upon the work initiated by Herman Muller in 1926 when he used X-rays to develop the first balancer chromosomes. Today, balancers are still invaluable to maintain mutations and transgenes but the arsenal of tools has expanded vastly and numerous new methods have been developed, many relying on the availability of the genome sequence and transposable elements. Forward genetic screens based on chemical mutagenesis or transposable elements have resulted in the unbiased identification of many novel players involved in processes probed by specific phenotypic assays. Reverse genetic approaches have relied on the availability of a carefully selected set of transposon insertions spread throughout the genome to allow the manipulation of the region in the vicinity of each insertion. Lastly, the ability to transform Drosophila with single copy transgenes using transposons or site-specific integration using the ΦC31 integrase has allowed numerous manipulations, including the ability to create and integrate genomic rescue constructs, generate duplications, RNAi knock-out technology, binary expression systems like the GAL4/UAS system as well as other methods. Here, we will discuss the most useful methodologies to interrogate the fruit fly genome in vivo focusing on chemical mutagenesis, transposons and transgenes. Genome engineering approaches based on nucleases and RNAi technology are discussed in following chapters.

Copyright © 2014 Elsevier Inc. All rights reserved.

KEYWORDS:

Chemical mutagens; Drosophila melanogaster; Enhancer bashing; Genetic screens; Genomic rescue; Mutation mapping; Overexpression; Transgenes; Transposon mutagenesis; Transposons

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