Tetracycline response element driven Cre causes ectopic recombinase activity independent of transactivator element
Objective: Tamoxifen is widely used for inducible Cre-LoxP systems but has several undesirable side effects for researchers investigating metabolism or energy balance, including weight loss, lipoatrophy, and drug incorporation into lipid stores. For this reason, we sought to determine whether a doxy...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-07-01
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| Series: | Molecular Metabolism |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2212877822000709 |
| _version_ | 1818253988877828096 |
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| author | Kenneth T. Lewis Lily R. Oles Ormond A. MacDougald |
| author_facet | Kenneth T. Lewis Lily R. Oles Ormond A. MacDougald |
| author_sort | Kenneth T. Lewis |
| collection | DOAJ |
| description | Objective: Tamoxifen is widely used for inducible Cre-LoxP systems but has several undesirable side effects for researchers investigating metabolism or energy balance, including weight loss, lipoatrophy, and drug incorporation into lipid stores. For this reason, we sought to determine whether a doxycycline-inducible system would be more advantageous for adipocyte-specific Cre mouse models, but serendipitously discovered widespread ectopic tetracycline response element Cre (TRE-Cre) recombinase activity. Methods: Adipocyte-specific tamoxifen- and doxycycline-inducible Cre mice were crossed to fluorescent Cre reporter mice and visualized by confocal microscopy to assess efficiency and background activity. TRE-Cre mice were crossed to stop-floxed diphtheria toxin mice to selectively ablate cells with background Cre activity. Results: Tamoxifen- and doxycycline-inducible systems performed similarly in adipose tissues, but ectopic Cre recombination was evident in numerous other cell types of the latter, most notably neurons. The source of ectopic Cre activity was isolated to the TRE-Cre transgene, driven by the pTet (tetO7) tetracycline-inducible promoter. Ablation of cells with ectopic recombination in mice led to stunted growth, diminished survival, and reduced brain mass. Conclusions: These results indicate that tamoxifen- and doxycycline-inducible adipocyte-specific Cre mouse models are similarly efficient, but the TRE-Cre component of the latter is inherently leaky. TRE-Cre background activity is especially pronounced in the brain and peripheral nerve fibers, and selective ablation of these cells impairs mouse development and survival. Caution should be taken when pairing TRE-Cre with floxed alleles that have defined roles in neural function, and additional controls should be included when using this model system. |
| first_indexed | 2024-12-12T16:48:49Z |
| format | Article |
| id | doaj.art-56487cc8b05f4fd79c9749331448ede2 |
| institution | Directory Open Access Journal |
| issn | 2212-8778 |
| language | English |
| last_indexed | 2024-12-12T16:48:49Z |
| publishDate | 2022-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Metabolism |
| spelling | doaj.art-56487cc8b05f4fd79c9749331448ede22022-12-22T00:18:25ZengElsevierMolecular Metabolism2212-87782022-07-0161101501Tetracycline response element driven Cre causes ectopic recombinase activity independent of transactivator elementKenneth T. Lewis0Lily R. Oles1Ormond A. MacDougald2University of Michigan Medical School, Department of Molecular & Integrative Physiology, Ann Arbor, MI, USAUniversity of Michigan Medical School, Department of Molecular & Integrative Physiology, Ann Arbor, MI, USAUniversity of Michigan Medical School, Department of Molecular & Integrative Physiology, Ann Arbor, MI, USA; University of Michigan Medical School, Department of Internal Medicine, Ann Arbor, MI, USA; Corresponding author. NCRC Building 25, Rm. 3686, 2800 Plymouth Rd., Ann Arbor, MI, 48105, USA.Objective: Tamoxifen is widely used for inducible Cre-LoxP systems but has several undesirable side effects for researchers investigating metabolism or energy balance, including weight loss, lipoatrophy, and drug incorporation into lipid stores. For this reason, we sought to determine whether a doxycycline-inducible system would be more advantageous for adipocyte-specific Cre mouse models, but serendipitously discovered widespread ectopic tetracycline response element Cre (TRE-Cre) recombinase activity. Methods: Adipocyte-specific tamoxifen- and doxycycline-inducible Cre mice were crossed to fluorescent Cre reporter mice and visualized by confocal microscopy to assess efficiency and background activity. TRE-Cre mice were crossed to stop-floxed diphtheria toxin mice to selectively ablate cells with background Cre activity. Results: Tamoxifen- and doxycycline-inducible systems performed similarly in adipose tissues, but ectopic Cre recombination was evident in numerous other cell types of the latter, most notably neurons. The source of ectopic Cre activity was isolated to the TRE-Cre transgene, driven by the pTet (tetO7) tetracycline-inducible promoter. Ablation of cells with ectopic recombination in mice led to stunted growth, diminished survival, and reduced brain mass. Conclusions: These results indicate that tamoxifen- and doxycycline-inducible adipocyte-specific Cre mouse models are similarly efficient, but the TRE-Cre component of the latter is inherently leaky. TRE-Cre background activity is especially pronounced in the brain and peripheral nerve fibers, and selective ablation of these cells impairs mouse development and survival. Caution should be taken when pairing TRE-Cre with floxed alleles that have defined roles in neural function, and additional controls should be included when using this model system.http://www.sciencedirect.com/science/article/pii/S2212877822000709Ectopic Cre activityTetracycline response elementpTetTRE-CretetO7-CreAdipoChaser |
| spellingShingle | Kenneth T. Lewis Lily R. Oles Ormond A. MacDougald Tetracycline response element driven Cre causes ectopic recombinase activity independent of transactivator element Molecular Metabolism Ectopic Cre activity Tetracycline response element pTet TRE-Cre tetO7-Cre AdipoChaser |
| title | Tetracycline response element driven Cre causes ectopic recombinase activity independent of transactivator element |
| title_full | Tetracycline response element driven Cre causes ectopic recombinase activity independent of transactivator element |
| title_fullStr | Tetracycline response element driven Cre causes ectopic recombinase activity independent of transactivator element |
| title_full_unstemmed | Tetracycline response element driven Cre causes ectopic recombinase activity independent of transactivator element |
| title_short | Tetracycline response element driven Cre causes ectopic recombinase activity independent of transactivator element |
| title_sort | tetracycline response element driven cre causes ectopic recombinase activity independent of transactivator element |
| topic | Ectopic Cre activity Tetracycline response element pTet TRE-Cre tetO7-Cre AdipoChaser |
| url | http://www.sciencedirect.com/science/article/pii/S2212877822000709 |
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