Abstract
Site-specific recombination systems, such as Cre-lox from bacteriophage P1, have become very important tools for plant genome engineering. In many cases a constitutive promoter is used to express the recombinase gene. However, for certain research and commercial applications constitutive Cre-mediated recombination may not be desirable. We have evaluated the potential of seven different germline promoter:cre fusions to remove a stably integrated lox cassette through Cre-mediated recombination in Arabidopsis thaliana. We monitored the functionality of each promoter in the germline of primary transformants by analyzing the presence of the recombined lox cassette in T2 progeny. The selected germline promoters are involved in different developmental cues, including early stem cell identity (CLAVATA3), flower meristem identity (LEAFY, APETALA1), floral organ identity (AGAMOUS), and meiosis (SOLO DANCERS, DMC1, SWITCH1). For five out of these seven promoters we were able to show that efficient Cre-mediated recombination does, indeed, occur and that the recombination takes place at some point during germline development. Furthermore, a recombination efficiency of 100% is obtained when Cre-expression is regulated by the CLAVATA3 promoter. In addition, with these promoters, we observe much less variation in recombination frequency than previously reported for the 35S promoter. For these reasons, we believe that germline-specific Cre-lox recombination provides an additional tool to the site-specific recombination technology in plants.
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Acknowledgments
The authors would like to acknowledge Dr. Sylvie De Buck for providing the FK24 line. Part of this research is funded by the Institute for Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen) through research project IWT-010067 and a predoctoral fellowship to FVE and by Research Foundation-Flanders (FWO; research project G.0067.09).
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Communicated by R. Schmidt.
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Van Ex, F., Verweire, D., Claeys, M. et al. Evaluation of seven promoters to achieve germline directed Cre-lox recombination in Arabidopsis thaliana . Plant Cell Rep 28, 1509–1520 (2009). https://doi.org/10.1007/s00299-009-0750-y
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DOI: https://doi.org/10.1007/s00299-009-0750-y