In-utero electroporation was performed as reported previously (García-Moreno et al. 2014 (link)) with modifications. Time-mated pregnant dames were administered Vetergesic intraperitoneally (0.08 mg/kg) for analgesia prior to anesthesia with Isoflurane. A midline laparotomy was undertaken and uterine horns exposed gently. DNA (mixed with Fast Green, Sigma) was injected into the lateral ventricles using pulled glass capillaries (Harvard Apparatus). Square pulses (at 30–35 V, 5 × 50 ms with 950 ms resting periods) were delivered using tweezer electrodes (CUY650P3, Nepagene) connected to a square pulse generator (Electro Square Porator, Harvard Apparatus). Warm PBS was used to keep the embryo sacs at near body temperature along with a warm blanket.
Following electroporation, the uterine horns were gently placed back in the abdominal cavity. Surgical incisions were sutured (Vicryl 4/0, Ethicon) and the skin held together using surgical clips. Animals were allowed to recover after removal of anesthesia and kept on heating pads until they regained movement.
Following electroporation, the uterine horns were gently placed back in the abdominal cavity. Surgical incisions were sutured (Vicryl 4/0, Ethicon) and the skin held together using surgical clips. Animals were allowed to recover after removal of anesthesia and kept on heating pads until they regained movement.
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