Construction and Detection of Fluorescent, Germline Transgenic Zebrafish

Zebrafish transgenesis was first demonstrated over 10 yr ago; however, the intensity of technological development has significantly increased in the past few years with the popularity of the zebrafish model. By transgenesis we are referring to germline transgenesis—the introduction of new genetic material into the germline, such that it is passed down to progeny and inherited in a Mendelian manner. This chapter describes the procedures used to produce transgenic zebrafish expressing fluorescent reporter genes via DNA microinjection. These procedures were modified from those developed by members of this laboratory during the establishment of the transgenic mouse facility at Duke Comprehensive Cancer Center. This chapter aims to aid others in efficiently producing transgenic zebrafish.