A Multiplex Gene Expression Assay for Direct Measurement of RNA Transcripts in Crude Lysates of the Nematode Caenorhabditis elegans Used as a Bioanalytical Tool

Gene expression profiling in Caenorhabditis elegans has been demonstrated to be a potential bioanalytical tool to detect the toxic potency of environmental contaminants. The RNA transcripts of genes responding to toxic exposure can be used as biomarkers for detecting these toxins. For routine applic...

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Veröffentlicht in:Environmental toxicology and chemistry 2023-01, Vol.42 (1), p.130-142
Hauptverfasser: Karengera, Antoine, Bao, Cong, Bovee, Toine F. H., Dinkla, Inez J. T., Murk, Albertinka J.
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container_title Environmental toxicology and chemistry
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creator Karengera, Antoine
Bao, Cong
Bovee, Toine F. H.
Dinkla, Inez J. T.
Murk, Albertinka J.
description Gene expression profiling in Caenorhabditis elegans has been demonstrated to be a potential bioanalytical tool to detect the toxic potency of environmental contaminants. The RNA transcripts of genes responding to toxic exposure can be used as biomarkers for detecting these toxins. For routine application in environmental quality monitoring, an easy‐to‐use multiplex assay is required to reliably quantify expression levels of these biomarkers. In the present study, a bead‐based assay was developed to fingerprint gene expression in C. elegans by quantitating messenger RNAs (mRNAs) of multiple target genes directly from crude nematode lysates, circumventing RNA extraction and purification steps. The assay uses signal amplification rather than target amplification for direct measurement of toxin‐induced RNA transcripts. Using a 50‐gene panel, the expression changes of four candidate reference genes and 46 target mRNAs for various contaminants and wastewaters were successfully measured, and the expression profiles indicated the type of toxin present. Moreover, the multiplex assay response was in line with previous results obtained with more time‐consuming reverse‐transcription quantitative polymerase chain reaction and microarray analyses. In addition, the transcriptomic profiles of nematodes exposed to wastewater samples and extracts prepared from tissues of swimming crabs were evaluated. The profiles indicated the presence of organic pollutants. The present study illustrates the successful development of a multiplex fluorescent bead–based approach using nematode C. elegans crude lysates for gene expression profiling of target RNAs. This method can be used to routinely fingerprint the presence of toxic contaminants in environmental samples and to identify the most biologically active fraction of the contaminant mixture in a toxicity identification and evaluation approach. Environ Toxicol Chem 2023;42:130–142. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.
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subjects Amplification
Animals
Assaying
Biological activity
Biomarkers
Caenorhabditis elegans
Caenorhabditis elegans - genetics
Contaminants
Crustaceans
DNA microarrays
Environmental monitoring
Environmental quality
Environmental Toxicology
Evaluation
Fingerprints
Fluorescence
Gene expression
Gene Expression Profiling
Genes
Lysates
multiplex assay
Multiplexing
nematode bioassay
Nematodes
Pollutants
Polymerase chain reaction
Ribonucleic acid
RNA
RNA, Messenger - metabolism
Swimming
Toxicity
Toxicology
Toxins
Transcriptome
Transcriptomics
Wastewater
water quality
Water sampling
title A Multiplex Gene Expression Assay for Direct Measurement of RNA Transcripts in Crude Lysates of the Nematode Caenorhabditis elegans Used as a Bioanalytical Tool
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