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. 2018 Apr 3;8:5435. doi: 10.1038/s41598-018-23810-8

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© The Author(s) 2018

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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Generation of ROSA26/CAG-floxed STOP-ChRFR(C167A)-Venus BAC actuator/reporter rats. (A) The transgene construct, which was inserted into the mouse ROSA26 BAC clone, comprised the CAG promoter, a cassette for the neomycin resistance gene with four STOP signal repeats (pA) flanked by loxP sites, and a sequence containing the ChRFR-C167A open reading frame. After Cre-mediated recombination, ChRFR(C167A)-Venus expression was under the regulation of the generic CAG promoter. (B) PCR analysis of genomic DNA. The 726-bp band, which is recognized by the designed primer sets targeting the PGK-neo cassette sequence between two loxP sequences, was present in the ChRFR(C167A)-Venus reporter rats but not in the wild-type rats. A 100-bp DNA ladder was used as marker (M). A full uncropped image is available as Supplementary Figure S1A.