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. 2020 Oct 20;147(2):361–371. doi: 10.1007/s00432-020-03428-y

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Gene cassettes integrated into the genome of MDA-MB-231 breast cancer cells. Scheme of two cassettes, which were transduced into MDA-MB-231 breast cancer cells. The upper cassette (transduced initially) contains: hPGK (human phosphoglycerate kinase promoter); tTA (tetracycline-controlled transactivator); IRES (internal ribosome entry site); eGFP (enhanced green fluorescent protein). The lower cassettes contains luciferase (firefly luciferase); P_tet-bi (bidirectional tet-regulated promoter); Cherry (red fluorescent protein mCherry); FRT and F3 (wild-type and mutant Flp—recombinase target sites); miRNA—artificial microRNA targeting specifically ITGB3 mRNA. The left bottom part illustrates the conditions in absence of doxycycline: tTA binds and activates the bidirectional promoter P_tet-bi, which leads to expression of firefly luciferase, red fluorescent protein mCherry and of the miRNA targeting ITGB3 mRNA. The right bottom part shows the consequences of doxycycline presence: tTA cannot bind to its respective promoter, it remains inactive and the expression of the subsequent genes is switched off