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High-throughput capturing and characterization of mutations in essential genes of Caenorhabditis elegans

BACKGROUND: Essential genes are critical for the development of all organisms and are associated with many human diseases. These genes have been a difficult category to study prior to the availability of balanced lethal strains. Despite the power of targeted mutagenesis, there are limitations in ide...

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Autores principales: Chu, Jeffrey Shih-Chieh, Chua, Shu-Yi, Wong, Kathy, Davison, Ann Marie, Johnsen, Robert, Baillie, David L, Rose, Ann M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4039747/
https://www.ncbi.nlm.nih.gov/pubmed/24884423
http://dx.doi.org/10.1186/1471-2164-15-361
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author Chu, Jeffrey Shih-Chieh
Chua, Shu-Yi
Wong, Kathy
Davison, Ann Marie
Johnsen, Robert
Baillie, David L
Rose, Ann M
author_facet Chu, Jeffrey Shih-Chieh
Chua, Shu-Yi
Wong, Kathy
Davison, Ann Marie
Johnsen, Robert
Baillie, David L
Rose, Ann M
author_sort Chu, Jeffrey Shih-Chieh
collection PubMed
description BACKGROUND: Essential genes are critical for the development of all organisms and are associated with many human diseases. These genes have been a difficult category to study prior to the availability of balanced lethal strains. Despite the power of targeted mutagenesis, there are limitations in identifying mutations in essential genes. In this paper, we describe the identification of coding regions for essential genes mutated using forward genetic screens in Caenorhabditis elegans. The lethal mutations described here were isolated and maintained by a wild-type allele on a rescuing duplication. RESULTS: We applied whole genome sequencing to identify the causative molecular lesion resulting in lethality in existing C. elegans mutant strains. These strains are balanced and can be easily maintained for subsequent characterization. Our method can be effectively used to analyze mutations in a large number of essential genes. We describe here the identification of 64 essential genes in a region of chromosome I covered by the duplication sDp2. Of these, 42 are nonsense mutations, six are splice signal mutations, one deletion, and 15 are non-synonymous mutations. Many of the essential genes in this region function in cell cycle, transcriptional regulation, and RNA processing. CONCLUSIONS: The essential genes identified here are represented by mutant strains, many of which have more than one mutant allele. The genetic resource can be utilized to further our understanding of essential gene function and will be applicable to the study of C. elegans development, conserved cellular function, and ultimately lead to improved human health. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2164-15-361) contains supplementary material, which is available to authorized users.
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spelling pubmed-40397472014-06-06 High-throughput capturing and characterization of mutations in essential genes of Caenorhabditis elegans Chu, Jeffrey Shih-Chieh Chua, Shu-Yi Wong, Kathy Davison, Ann Marie Johnsen, Robert Baillie, David L Rose, Ann M BMC Genomics Research Article BACKGROUND: Essential genes are critical for the development of all organisms and are associated with many human diseases. These genes have been a difficult category to study prior to the availability of balanced lethal strains. Despite the power of targeted mutagenesis, there are limitations in identifying mutations in essential genes. In this paper, we describe the identification of coding regions for essential genes mutated using forward genetic screens in Caenorhabditis elegans. The lethal mutations described here were isolated and maintained by a wild-type allele on a rescuing duplication. RESULTS: We applied whole genome sequencing to identify the causative molecular lesion resulting in lethality in existing C. elegans mutant strains. These strains are balanced and can be easily maintained for subsequent characterization. Our method can be effectively used to analyze mutations in a large number of essential genes. We describe here the identification of 64 essential genes in a region of chromosome I covered by the duplication sDp2. Of these, 42 are nonsense mutations, six are splice signal mutations, one deletion, and 15 are non-synonymous mutations. Many of the essential genes in this region function in cell cycle, transcriptional regulation, and RNA processing. CONCLUSIONS: The essential genes identified here are represented by mutant strains, many of which have more than one mutant allele. The genetic resource can be utilized to further our understanding of essential gene function and will be applicable to the study of C. elegans development, conserved cellular function, and ultimately lead to improved human health. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2164-15-361) contains supplementary material, which is available to authorized users. BioMed Central 2014-05-12 /pmc/articles/PMC4039747/ /pubmed/24884423 http://dx.doi.org/10.1186/1471-2164-15-361 Text en © Chu et al.; licensee BioMed Central Ltd. 2014 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Chu, Jeffrey Shih-Chieh
Chua, Shu-Yi
Wong, Kathy
Davison, Ann Marie
Johnsen, Robert
Baillie, David L
Rose, Ann M
High-throughput capturing and characterization of mutations in essential genes of Caenorhabditis elegans
title High-throughput capturing and characterization of mutations in essential genes of Caenorhabditis elegans
title_full High-throughput capturing and characterization of mutations in essential genes of Caenorhabditis elegans
title_fullStr High-throughput capturing and characterization of mutations in essential genes of Caenorhabditis elegans
title_full_unstemmed High-throughput capturing and characterization of mutations in essential genes of Caenorhabditis elegans
title_short High-throughput capturing and characterization of mutations in essential genes of Caenorhabditis elegans
title_sort high-throughput capturing and characterization of mutations in essential genes of caenorhabditis elegans
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4039747/
https://www.ncbi.nlm.nih.gov/pubmed/24884423
http://dx.doi.org/10.1186/1471-2164-15-361
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