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. 2017 Jun 20;7:3881. doi: 10.1038/s41598-017-03984-3

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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The new transgenic parasite lines expressing DiCre recombinase excise floxed DNA sequences with >99% efficiency using rapamycin at lower concentrations and short treatment times. (a) Live GFP fluorescence assay of wild type and DiCre transgenic parasite lines transfected with the reporter plasmid pHH3-SP-loxPint-GFP-loxP and treated at the early ring stage with DMSO or rapamycin as indicated on top of the panels. Images were taken 40 h later. Size bars equal 10 μm. (b) Excision efficiency determined by flow cytometry. The four parasite lines transfected with the reporter plasmid were treated with DMSO or rapamycin as described above and the number of GFP positive schizonts was counted 38–40 h after treatment. Data correspond to the mean and standard deviation (s.d.) calculated from three independent experiments counting 100,000 cells each time.