Selection of Reliable Reference Genes in for Analysis of Nanotoxicity

ECU Author/Contributor (non-ECU co-authors, if there are any, appear on document)

Dongliang Chen (Creator)

Xiaoping Pan (Creator)

Michael A. Smith (Creator)

Baohong Zhang (Creator)

Yanqiong Zhang (Creator)

Institution

East Carolina University (ECU )

Web Site: http://www.ecu.edu/lib/

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 ?Ct 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.

Additional Information

Publication

Other

PLoS ONE; 7:3 p. 1-7

Language: English

Date: 2012

Keywords

Algorithms, Larvae, Gene expression, RNA extraction, Computer software, Nanoparticles

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