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Selection of Reliable Reference Genes in Caenorhabditis elegans for Analysis of Nanotoxicity

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  • Yanqiong Zhang
  • Dongliang Chen
  • Michael A Smith
  • Baohong Zhang
  • Xiaoping Pan

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.

Suggested Citation

  • Yanqiong Zhang & Dongliang Chen & Michael A Smith & Baohong Zhang & Xiaoping Pan, 2012. "Selection of Reliable Reference Genes in Caenorhabditis elegans for Analysis of Nanotoxicity," PLOS ONE, Public Library of Science, vol. 7(3), pages 1-7, March.
    • Handle: RePEc:plo:pone00:0031849
    DOI: 10.1371/journal.pone.0031849
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    Cited by:

    1. Hyo Sub Choi & Ajay Bhat & Marshall B. Howington & Megan L. Schaller & Rebecca L. Cox & Shijiao Huang & Safa Beydoun & Hillary A. Miller & Angela M. Tuckowski & Joy Mecano & Elizabeth S. Dean & Lindy , 2023. "FMO rewires metabolism to promote longevity through tryptophan and one carbon metabolism in C. elegans," Nature Communications, Nature, vol. 14(1), pages 1-16, December.
    2. Martín Bustelo & Martín A Bruno & César F Loidl & Manuel Rey-Funes & Harry W M Steinbusch & Antonio W D Gavilanes & D L A van den Hove, 2020. "Statistical differences resulting from selection of stable reference genes after hypoxia and hypothermia in the neonatal rat brain," PLOS ONE, Public Library of Science, vol. 15(5), pages 1-12, May.

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