In utero electroporation of mouse embryos
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Corresponding Organization : Hôpitaux Universitaires de Strasbourg
Other organizations : Université Paris Cité, Institut Cochin, Manchester Academic Health Science Centre, University of Manchester, University of Oxford, Wellcome Centre for Human Genetics, Centre Hospitalier Universitaire de Nantes, Centre Hospitalier Universitaire de Montpellier, Boston Children's Hospital, University of Liège, Université de Reims Champagne-Ardenne, Hôpital Maison Blanche, Centre Hospitalier Universitaire de Reims, University of Florence, Meyer Children's Hospital, Commissariat à l'Énergie Atomique et aux Énergies Alternatives, Genoscope, Institut des Maladies Génétiques Imagine, Research Institute of Molecular Pathology, Hôpital Cochin, Oxford University Hospitals NHS Trust, Hôpital Necker-Enfants Malades
Variable analysis
- In utero electroporation performed as described previously13 (link),40 (link) using Swiss mice (Janvier)
- Electroporated cells visualized using fluorescent Tomato protein
- Timed pregnant mice (E14.5)
- Uterine horns exposed
- Lateral ventricle of each embryo injected with Fast Green (2 μg/ml; Sigma) combined with a final concentration of 1µg/µl of DNA constructs
- Expression vector pCAGGS-Tomato systematically co-electroporated
- Electric pulses delivered at 50V for 50ms at 950ms intervals using a CUY21EDIT electroporator (Sonidel Ltd)
- Embryos or pups brains dissected and fixed in 4% PFA in PBS overnight
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