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

BMC Genomics

Ann M Rose²  David L Baillie¹  Robert Johnsen¹  Ann Marie Davison³  Kathy Wong²  Shu-Yi Chua¹  Jeffrey Shih-Chieh Chu² 

[1] Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, Canada;Department of Medical Genetics, University of British Columbia, Vancouver, Canada;Department of Biology, Kwantlen Polytechnic University, Surrey, Canada

关键词: Cell cycle;    C. elegans;    Lethal mutation;    Balanced mutation;    Essential genes;    Mutagenesis;    EMS;    Whole genome sequencing;   

Others  :  1217238
DOI  :  10.1186/1471-2164-15-361

 received in 2013-12-05, accepted in 2014-05-06,  发布年份 2014

【 摘 要 】

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.

【 授权许可】

   
2014 Chu et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

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