Comparison of the <i>Opn-CreER</i> and <i>Ck19-CreER</i> Drivers in Bile Ducts of Normal and Injured Mouse Livers
Inducible cyclization recombinase (Cre) transgenic mouse strains are powerful tools for cell lineage tracing and tissue-specific knockout experiments. However, low efficiency or leaky expression can be important pitfalls. Here, we compared the efficiency and specificity of two commonly used cholangi...
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MDPI AG
2019-04-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/8/4/380 |
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| author | Bram Lesaffer Elisabeth Verboven Leen Van Huffel Iván M. Moya Leo A. van Grunsven Isabelle A. Leclercq Frédéric P. Lemaigre Georg Halder |
| author_facet | Bram Lesaffer Elisabeth Verboven Leen Van Huffel Iván M. Moya Leo A. van Grunsven Isabelle A. Leclercq Frédéric P. Lemaigre Georg Halder |
| author_sort | Bram Lesaffer |
| collection | DOAJ |
| description | Inducible cyclization recombinase (Cre) transgenic mouse strains are powerful tools for cell lineage tracing and tissue-specific knockout experiments. However, low efficiency or leaky expression can be important pitfalls. Here, we compared the efficiency and specificity of two commonly used cholangiocyte-specific Cre drivers, the <i>Opn-iCreER<sup>T2</sup></i> and <i>Ck19-CreER<sup>T</sup></i> drivers, using a tdTomato reporter strain. We found that <i>Opn-iCreER<sup>T2</sup></i> triggered recombination of the tdTomato reporter in 99.9% of all cholangiocytes while <i>Ck19-CreER<sup>T</sup></i> only had 32% recombination efficiency after tamoxifen injection. In the absence of tamoxifen, recombination was also induced in 2% of cholangiocytes for the <i>Opn-iCreER<sup>T2</sup></i> driver and in 13% for the <i>Ck19-CreER<sup>T</sup></i> driver. For both drivers, Cre recombination was highly specific for cholangiocytes since recombination was rare in other liver cell types. Toxic liver injury ectopically activated <i>Opn-iCreER<sup>T2</sup></i> but not <i>Ck19-CreER<sup>T</sup></i> expression in hepatocytes. However, ectopic recombination in hepatocytes could be avoided by applying a three-day long wash-out period between tamoxifen treatment and toxin injection. Therefore, the <i>Opn-iCreER<sup>T2</sup></i> driver is best suited for the generation of mutant bile ducts, while the <i>Ck19-CreER<sup>T</sup></i> driver has near absolute specificity for bile duct cells and is therefore favorable for lineage tracing experiments. |
| first_indexed | 2024-03-12T08:02:07Z |
| format | Article |
| id | doaj.art-92621a6794e94aaf8c029bbd47c489fc |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-12T08:02:07Z |
| publishDate | 2019-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-92621a6794e94aaf8c029bbd47c489fc2023-09-02T19:47:27ZengMDPI AGCells2073-44092019-04-018438010.3390/cells8040380cells8040380Comparison of the <i>Opn-CreER</i> and <i>Ck19-CreER</i> Drivers in Bile Ducts of Normal and Injured Mouse LiversBram Lesaffer0Elisabeth Verboven1Leen Van Huffel2Iván M. Moya3Leo A. van Grunsven4Isabelle A. Leclercq5Frédéric P. Lemaigre6Georg Halder7VIB Center for Cancer Biology and KU Leuven Department of Oncology, University of Leuven, 3000 Leuven, BelgiumVIB Center for Cancer Biology and KU Leuven Department of Oncology, University of Leuven, 3000 Leuven, BelgiumVIB Center for Cancer Biology and KU Leuven Department of Oncology, University of Leuven, 3000 Leuven, BelgiumVIB Center for Cancer Biology and KU Leuven Department of Oncology, University of Leuven, 3000 Leuven, BelgiumLiver Cell Biology research group, Vrije Universiteit Brussel, 1090 Brussels, BelgiumLaboratory of Hepato-gastroenterology, Institut de Recherche Expérimentale et Clinique, Université catholique de Louvain, 1200 Brussels, BelgiumLiver and Pancreas Development Unit, de Duve Institute, Université catholique de Louvain, 1200 Brussels, BelgiumVIB Center for Cancer Biology and KU Leuven Department of Oncology, University of Leuven, 3000 Leuven, BelgiumInducible cyclization recombinase (Cre) transgenic mouse strains are powerful tools for cell lineage tracing and tissue-specific knockout experiments. However, low efficiency or leaky expression can be important pitfalls. Here, we compared the efficiency and specificity of two commonly used cholangiocyte-specific Cre drivers, the <i>Opn-iCreER<sup>T2</sup></i> and <i>Ck19-CreER<sup>T</sup></i> drivers, using a tdTomato reporter strain. We found that <i>Opn-iCreER<sup>T2</sup></i> triggered recombination of the tdTomato reporter in 99.9% of all cholangiocytes while <i>Ck19-CreER<sup>T</sup></i> only had 32% recombination efficiency after tamoxifen injection. In the absence of tamoxifen, recombination was also induced in 2% of cholangiocytes for the <i>Opn-iCreER<sup>T2</sup></i> driver and in 13% for the <i>Ck19-CreER<sup>T</sup></i> driver. For both drivers, Cre recombination was highly specific for cholangiocytes since recombination was rare in other liver cell types. Toxic liver injury ectopically activated <i>Opn-iCreER<sup>T2</sup></i> but not <i>Ck19-CreER<sup>T</sup></i> expression in hepatocytes. However, ectopic recombination in hepatocytes could be avoided by applying a three-day long wash-out period between tamoxifen treatment and toxin injection. Therefore, the <i>Opn-iCreER<sup>T2</sup></i> driver is best suited for the generation of mutant bile ducts, while the <i>Ck19-CreER<sup>T</sup></i> driver has near absolute specificity for bile duct cells and is therefore favorable for lineage tracing experiments.https://www.mdpi.com/2073-4409/8/4/380Crecholangiocytesbile duct cellsknockoutmouse liverlineage tracingOpnCk19 |
| spellingShingle | Bram Lesaffer Elisabeth Verboven Leen Van Huffel Iván M. Moya Leo A. van Grunsven Isabelle A. Leclercq Frédéric P. Lemaigre Georg Halder Comparison of the <i>Opn-CreER</i> and <i>Ck19-CreER</i> Drivers in Bile Ducts of Normal and Injured Mouse Livers Cells Cre cholangiocytes bile duct cells knockout mouse liver lineage tracing Opn Ck19 |
| title | Comparison of the <i>Opn-CreER</i> and <i>Ck19-CreER</i> Drivers in Bile Ducts of Normal and Injured Mouse Livers |
| title_full | Comparison of the <i>Opn-CreER</i> and <i>Ck19-CreER</i> Drivers in Bile Ducts of Normal and Injured Mouse Livers |
| title_fullStr | Comparison of the <i>Opn-CreER</i> and <i>Ck19-CreER</i> Drivers in Bile Ducts of Normal and Injured Mouse Livers |
| title_full_unstemmed | Comparison of the <i>Opn-CreER</i> and <i>Ck19-CreER</i> Drivers in Bile Ducts of Normal and Injured Mouse Livers |
| title_short | Comparison of the <i>Opn-CreER</i> and <i>Ck19-CreER</i> Drivers in Bile Ducts of Normal and Injured Mouse Livers |
| title_sort | comparison of the i opn creer i and i ck19 creer i drivers in bile ducts of normal and injured mouse livers |
| topic | Cre cholangiocytes bile duct cells knockout mouse liver lineage tracing Opn Ck19 |
| url | https://www.mdpi.com/2073-4409/8/4/380 |
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