The use of in ovo electroporation for the rapid analysis of neural-specific murine enhancers

The identification and characterization of DNA sequences necessary for prop er gene expression have provided insights into gene regulation and generate d tools useful for further experimentation. Studies of developmentally regu lated genes have demonstrated how transcription factors interact at enhance rs to generate restricted patterns of expression during embryogenesis, In v ertebrates, the pursuit of such studies has relied on the generation of tra nsgenic mice and thus has been limited by the time and expense required gen erating and characterizing these mice. The recently developed technique of in ovo electroporation allows the rapid introduction of exogenous DNA into developing chicken embryos. Here we have used this technique to introduce D NA containing murine enhancer/reporter constructs into cells of the chicken neural tube, resulting in appropriate expression of the reporter. This tec hnique has the potential to greatly reduce the effort involved in the study of vertebrate enhancers. Furthermore, we have characterized factors such a s timing of electroporation, concentration of DNA, and choice of basal prom oters and found that they can influence the degree to which expression of e nhancer constructs reflects endogenous gene expression, (C) 2001 Wiley-Liss , Inc.