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. 2019 Oct 22;10:4801. doi: 10.1038/s41467-019-12794-2

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© The Author(s) 2019

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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Fig. 2 — Construction of sensory switch circuit. a Functional analysis of AFP promoter (III) in different cell lines by transient transfection. The constitutive CAG promoter was used as a positive control. b Functional analysis of microRNAs in indicated cell lines. Four tandem repeats of microRNA binding sites were inserted into the 3′-UTR of EYFP reporter gene. The CMV-driven EBFP was used as an internal control in transient transfection experiments. c Characterization of sensory switch circuit in HEK293 cells by transient transfection. LacO binding site of LacI repressor, tetO binding site of tetR: Krab repressor, L self-cleavage 2A linker, FF4 shRNA-FF4, FF5 shRNA-FF5, the right bottom panel shows representative flow cytometry scatter plots measured 48 h after transfection. d Hierarchical assembly of sensory switch circuit and loading into adenoviral vector. E in blue circle, Esp3I; B in orange circle, BsaI; L in box, self-cleavage 2A linker; GI1~4 and GII1~4, different overhangs released by Esp3I or BsaI; ccdB, ccdB toxin coding gene; RSL and RSR left and right Gateway recombination site, mbs microRNA binding sites, Kana kanamycin resistance gene, Tet tetracycline resistance gene, Amp ampicillin resistance gene, PacI PacI cutting site, ITR inverted terminal repeat, PS packaging signal. a~c Each data point shows mean ± s.d. of EYFP or EBFP from three independent replicates. Source data are provided as a Source Data file