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Large-scale identification of chemically induced mutations in Drosophila melanogaster

Forward genetic screens using chemical mutagens have been successful in defining the function of thousands of genes in eukaryotic model organisms. The main drawback of this strategy is the time-consuming identification of the molecular lesions causative of the phenotypes of interest. With whole-geno...

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Autores principales: Haelterman, Nele A., Jiang, Lichun, Li, Yumei, Bayat, Vafa, Sandoval, Hector, Ugur, Berrak, Tan, Kai Li, Zhang, Ke, Bei, Danqing, Xiong, Bo, Charng, Wu-Lin, Busby, Theodore, Jawaid, Adeel, David, Gabriela, Jaiswal, Manish, Venken, Koen J.T., Yamamoto, Shinya, Chen, Rui, Bellen, Hugo J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4199363/
https://www.ncbi.nlm.nih.gov/pubmed/25258387
http://dx.doi.org/10.1101/gr.174615.114
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author Haelterman, Nele A.
Jiang, Lichun
Li, Yumei
Bayat, Vafa
Sandoval, Hector
Ugur, Berrak
Tan, Kai Li
Zhang, Ke
Bei, Danqing
Xiong, Bo
Charng, Wu-Lin
Busby, Theodore
Jawaid, Adeel
David, Gabriela
Jaiswal, Manish
Venken, Koen J.T.
Yamamoto, Shinya
Chen, Rui
Bellen, Hugo J.
author_facet Haelterman, Nele A.
Jiang, Lichun
Li, Yumei
Bayat, Vafa
Sandoval, Hector
Ugur, Berrak
Tan, Kai Li
Zhang, Ke
Bei, Danqing
Xiong, Bo
Charng, Wu-Lin
Busby, Theodore
Jawaid, Adeel
David, Gabriela
Jaiswal, Manish
Venken, Koen J.T.
Yamamoto, Shinya
Chen, Rui
Bellen, Hugo J.
author_sort Haelterman, Nele A.
collection PubMed
description Forward genetic screens using chemical mutagens have been successful in defining the function of thousands of genes in eukaryotic model organisms. The main drawback of this strategy is the time-consuming identification of the molecular lesions causative of the phenotypes of interest. With whole-genome sequencing (WGS), it is now possible to sequence hundreds of strains, but determining which mutations are causative among thousands of polymorphisms remains challenging. We have sequenced 394 mutant strains, generated in a chemical mutagenesis screen, for essential genes on the Drosophila X chromosome and describe strategies to reduce the number of candidate mutations from an average of ∼3500 to 35 single-nucleotide variants per chromosome. By combining WGS with a rough mapping method based on large duplications, we were able to map 274 (∼70%) mutations. We show that these mutations are causative, using small 80-kb duplications that rescue lethality. Hence, our findings demonstrate that combining rough mapping with WGS dramatically expands the toolkit necessary for assigning function to genes.
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spelling pubmed-41993632015-04-01 Large-scale identification of chemically induced mutations in Drosophila melanogaster Haelterman, Nele A. Jiang, Lichun Li, Yumei Bayat, Vafa Sandoval, Hector Ugur, Berrak Tan, Kai Li Zhang, Ke Bei, Danqing Xiong, Bo Charng, Wu-Lin Busby, Theodore Jawaid, Adeel David, Gabriela Jaiswal, Manish Venken, Koen J.T. Yamamoto, Shinya Chen, Rui Bellen, Hugo J. Genome Res Method Forward genetic screens using chemical mutagens have been successful in defining the function of thousands of genes in eukaryotic model organisms. The main drawback of this strategy is the time-consuming identification of the molecular lesions causative of the phenotypes of interest. With whole-genome sequencing (WGS), it is now possible to sequence hundreds of strains, but determining which mutations are causative among thousands of polymorphisms remains challenging. We have sequenced 394 mutant strains, generated in a chemical mutagenesis screen, for essential genes on the Drosophila X chromosome and describe strategies to reduce the number of candidate mutations from an average of ∼3500 to 35 single-nucleotide variants per chromosome. By combining WGS with a rough mapping method based on large duplications, we were able to map 274 (∼70%) mutations. We show that these mutations are causative, using small 80-kb duplications that rescue lethality. Hence, our findings demonstrate that combining rough mapping with WGS dramatically expands the toolkit necessary for assigning function to genes. Cold Spring Harbor Laboratory Press 2014-10 /pmc/articles/PMC4199363/ /pubmed/25258387 http://dx.doi.org/10.1101/gr.174615.114 Text en © 2014 Haelterman et al.; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genome.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Method
Haelterman, Nele A.
Jiang, Lichun
Li, Yumei
Bayat, Vafa
Sandoval, Hector
Ugur, Berrak
Tan, Kai Li
Zhang, Ke
Bei, Danqing
Xiong, Bo
Charng, Wu-Lin
Busby, Theodore
Jawaid, Adeel
David, Gabriela
Jaiswal, Manish
Venken, Koen J.T.
Yamamoto, Shinya
Chen, Rui
Bellen, Hugo J.
Large-scale identification of chemically induced mutations in Drosophila melanogaster
title Large-scale identification of chemically induced mutations in Drosophila melanogaster
title_full Large-scale identification of chemically induced mutations in Drosophila melanogaster
title_fullStr Large-scale identification of chemically induced mutations in Drosophila melanogaster
title_full_unstemmed Large-scale identification of chemically induced mutations in Drosophila melanogaster
title_short Large-scale identification of chemically induced mutations in Drosophila melanogaster
title_sort large-scale identification of chemically induced mutations in drosophila melanogaster
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4199363/
https://www.ncbi.nlm.nih.gov/pubmed/25258387
http://dx.doi.org/10.1101/gr.174615.114
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