Abstract
Transgenic mutation assays are used to identify and characterize genotoxic hazards and for determining the mode of action for carcinogens. The three most popular transgenic mutational models are Big Blue® (rats or mice), Muta™ mouse (mice), and gpt-delta (rats or mice). The Big Blue® and Muta™ mouse models use the cII gene as a reporter of mutation whereas gpt-delta rodents use the gpt gene and the red/gam genes (Spi− selection) as mutation reporter genes. Here we describe methodology for conducting mutation assays with these transgenes. Transgenes recovered from tissue DNA are packaged into infectious lambda phage, bacteria are infected with the phage, and cII-mutant and Spi− plaques and gpt-mutant colonies are isolated using selective conditions and quantified. Selected mutants can be further analyzed for identification of small sequence alterations in the cII and gpt genes and large deletions at the Spi− locus.
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Lambert IB, Singer TM, Boucher SE et al (2005) Detailed review of transgenic rodent mutation assays. Mutat Res 590:1–280
Organization for Economic Cooperation and Development (OECD) (2011). OECD guideline for the testing of chemicals; transgenic rodent somatic and germ cell gene mutation assay. Available at: http://www.oecd-ilibrary.org/docserver/download/fulltext/9748801e.pdf?expires=1342710912&id=id&accname=freeContent&checksum=06A7713CC9C6E1B7EEFCC61F2565CD7
Pozniak A, Muller L, Salgo M et al (2009) Elevated EMS in nelfinavir mesylate (Viracept, Roche): overview. AIDS Res Ther 6:18–23
Boverhof DR, Chamberlain MP, Elcombe CR et al (2011) Transgenic animal models in toxicology: historical perspectives and future outlook. Toxicol Sci 121:207–233
Heddle JA, Martus HJ, Douglas GR (2003) Treatment and sampling protocols for transgenic mutation assays. Environ Mol Mutagen 41:1–6
Thybaud V, Dean S, Nohmi T et al (2003) In vivo transgenic mutation assays. Mutat Res 540:141–151
Acknowledgement
We are grateful to our colleague, Vasily N. Dobrovolsky, for help in making the figure.
Disclaimer: The guidelines for conducting the transgenic assays described in this chapter are not necessarily endorsed by the US Food and Drug Administration.
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Manjanatha, M.G., Cao, X., Shelton, S.D., Mittelstaedt, R.A., Heflich, R.H. (2013). In Vivo cII, gpt, and Spi− Gene Mutation Assays in Transgenic Mice and Rats. In: Dhawan, A., Bajpayee, M. (eds) Genotoxicity Assessment. Methods in Molecular Biology, vol 1044. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-529-3_5
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DOI: https://doi.org/10.1007/978-1-62703-529-3_5
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-528-6
Online ISBN: 978-1-62703-529-3
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