Abstract
Transgene silencing is often associated with multicopy integrations, which occur frequently during plant transformation. Transgene expression can be restored in a number of multicopy loci by converting them to single copy. This chapter describes a plant transformation protocol based on use of the Cre-lox system, which allows conversion of a multicopy transgene locus into single copy. The strategy is based on designing a transformation vector with lox sites, developing transgenic lines, and introducing Cre activity to initiate Cre-lox recombination, which leads to the simplification of a multicopy locus to a single- or low-copy state. This method is compatible with both gene gun and Agrobacterium-mediated gene delivery and should be particularly useful for crops that are difficult to transform.
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Acknowledgments
Research support from the USDA-NIFA (2006-33120-17718; 2010-33522-21715) and the Arkansas Bioscience Institute to V. Srivastava, and from the Chinese Ministry of Agriculture (2010ZX08010-001) to D. W. Ow are gratefully acknowledged.
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Srivastava, V., Ow, D.W. (2015). Simplifying Transgene Locus Structure Through Cre-lox Recombination. In: Mysore, K., Senthil-Kumar, M. (eds) Plant Gene Silencing. Methods in Molecular Biology, vol 1287. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2453-0_6
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DOI: https://doi.org/10.1007/978-1-4939-2453-0_6
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