Abstract
Herein we demonstrate with PCR, immunodetection and reporter gene approaches that the widely used human Glial Fibrillary Acidic Protein (hGFAP)-Cre transgene exhibits spontaneous germ-line recombination activity in leading to deletion in brain, heart and tail tissue with high frequency. The ectopic activity of hGFAP-Cre requires a rigorous control. We likewise observed that a second widely used nestin-Cre transgene shows germ-line deletion. Here we describe procedures to identify mice with germ-line recombination mediated by the hGFAP-Cre and nestin-Cre transgenes. Such control is essential to avoid pleiotropic effects due to germ-line deletion of loxP-flanked target genes and to maintain the CNS-restricted deletion status in transgenic mouse colonies.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cerebral Cortex / metabolism
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Connexin 43 / genetics
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Female
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Gene Deletion
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Gene Knockout Techniques
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Gene Order
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Germ Cells / metabolism*
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Glial Fibrillary Acidic Protein / genetics*
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Glial Fibrillary Acidic Protein / metabolism
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Hippocampus / metabolism
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Homologous Recombination*
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Humans
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Immunohistochemistry
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Integrases / genetics*
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Integrases / metabolism*
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Male
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Mice
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Mice, Knockout
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Mice, Transgenic
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Myocardium / metabolism
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Nestin / genetics*
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Transgenes
Substances
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Connexin 43
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Glial Fibrillary Acidic Protein
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Nestin
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Cre recombinase
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Integrases
Grants and funding
This work was supported by grants of the German Research Foundation SFB/TR3, C1 and C9; SPP1172 SE 774/3 (to CS); SFB/TR3, N01 and C9; SPP1172 TH1350/1-1 (to MT) and of the European Community FP7-202167 (to CS and MT). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.