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

Authors: Chu, Jeffrey S; Chua, Shu-Yi; Wong, Kathy; Davison, Ann M; Johnsen, Robert; Baillie, David L; Rose, Ann M;

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

Abstract

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.

Keywords

Lethal mutation, 570, Transcription, Genetic, Cell cycle, Essential genes, Chromosomes, Open Reading Frames, Balanced mutation, Genetics, Animals, Caenorhabditis elegans, Caenorhabditis elegans Proteins, Alleles, Cytokinesis, Genes, Essential, Whole Genome Sequencing, Sequence Analysis, RNA, EMS, High-Throughput Nucleotide Sequencing, Mutagenesis, Whole genome sequencing, Mutation, C. elegans, RNA, RNA Interference, Biotechnology, Research Article

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    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
16
Top 10%
Average
Top 10%
Green
gold