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Selection of reliable reference genes in Caenorhabditis elegans for analysis of nanotoxicity.
- Source :
-
PloS one [PLoS One] 2012; Vol. 7 (3), pp. e31849. Date of Electronic Publication: 2012 Mar 15. - Publication Year :
- 2012
-
Abstract
- Despite rapid development and application of a wide range of manufactured metal oxide nanoparticles (NPs), the understanding of potential risks of using NPs is less completed, especially at the molecular level. The nematode Caenorhabditis elegans (C.elegans) has been emerging as an environmental model to study the molecular mechanism of environmental contaminations, using standard genetic tools such as the real-time quantitative PCR (RT-qPCR). The most important factor that may affect the accuracy of RT-qPCR is to choose appropriate genes for normalization. In this study, we selected 13 reference gene candidates (act-1, cdc-42, pmp-3, eif-3.C, actin, act-2, csq-1, Y45F10D.4, tba-1, mdh-1, ama-1, F35G12.2, and rbd-1) to test their expression stability under different doses of nano-copper oxide (CuO 0, 1, 10, and 50 µg/mL) using RT-qPCR. Four algorithms, geNorm, NormFinder, BestKeeper, and the comparative ΔCt method, were employed to evaluate these 13 candidates expressions. As a result, tba-1, Y45F10D.4 and pmp-3 were the most reliable, which may be used as reference genes in future study of nanoparticle-induced genetic response using C.elegans.
- Subjects :
- Animals
Animals, Genetically Modified
Base Sequence
Copper toxicity
DNA Primers genetics
DNA, Helminth genetics
Gene Expression
Models, Animal
Nanotechnology
Reverse Transcriptase Polymerase Chain Reaction
Caenorhabditis elegans drug effects
Caenorhabditis elegans genetics
Genes, Helminth drug effects
Metal Nanoparticles toxicity
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 7
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 22438870
- Full Text :
- https://doi.org/10.1371/journal.pone.0031849