Different Gene Transfer Methods at the Very Early, Early, Late and Whole Embryonic Stages in Chicken
New technologies in gene transfer combined with experimental embryology make the chicken embryo an excellent model system for gene function studies. The techniques of in ovo electroporation, in vitro culture for ex ovo electroporation and retrovirus-mediated gene transfer have already been fully developed in chicken. Yet to our knowledge, there are no definite descriptions on the features and application scopes of these techniques. The survival rates of different in vitro culture methods were compared and the EGFP expression areas of different gene transfer techniques were explored. It was that the optimal timings of removing embryo for EC culture and Petri dish system was at El.5 and E2.5, respectively; and optimal timing of injecting retrovirus is at E0. Results indicated that the EC culture, in ovo electroporation, the Petri dish system and retrovirus-mediated method are, respectively, suitable for the very early, early, late and whole embryonic stages in chicken. Comparison of different gene transfer methods and establishment of optimal timings are expected to provide a better choice of the efficient method for a particular experiment.
KeywordsChicken embryo gene transfer in ovo electroporation in vitro culture retrovirus
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We are grateful to Dr. Yoshiko Takahashi (Nara Institute of Science and Technology, Japan) for the kind gifts of pBI-EGFP and pCAGGS-rtTA2S-M2, and also to Professor Cliff Tabin (Harvard Medical School, USA) for the kind gift of RCASBP.B viral vector. This work was supported by the National Natural Science Foundation of China (30800781 and 31072022) and New faculty funding of Ministry of Education of P. R. China (200805041054).
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