piggyBac transposition into primordial germ cells is an efficient tool for transgenesis in chickens.
Transgenic birds embody one of the most potent and exciting research tools in biotechnology for agriculture, medicine, and model animals. To date, retrovirus- or lentivirus-mediated transgenesis has been established in chickens and quail. However, despite having a valid technique for viral transduction to achieve transgenic birds, many obstacles exist for practical applications because of relatively low and variable rates of germ-line transmission and transgenic offspring showing transgene silencing, as well as safety issues related to viral vector use. Thus, the generation of transgenic poultry by nonviral integration is a prerequisite for the introduction of biotechnology to practical applications. Herein, we show that a germ-line-competent chicken primordial germ-cell (PGC) line was established with high efficiency of transmission to offspring and that piggyBac transposition into PGCs improved the efficiency of transgenic chicken production and led to high-level transgene expression. GFP transgene-expressing donor PGC-transferred recipient chickens produced donor-derived progenies, and the germ-line transmission efficiency of donor PGCs was 95.2% on average. Subsequently, half of the donor-derived offspring (52.2%) were transgenic chicks because GFP-expressing donor PGCs, in which a transgene was inserted into one chromosome 20, were heterozygous. In all of the transgenic chickens, GFP expression was constant and strong, regardless of age. Our results demonstrate that piggyBac transposition into the chicken PGC line could be the surest way to generate transgenic chickens safely for practical applications.
World Class University Biomodulation Major, Department of Agricultural Biotechnology, Research Institute for Agriculture and Life Sciences, College of Agriculture and Life Sciences,  Seoul National University, Seoul 151-921,  Korea.
Tae Sub
TS
Park
Jae Yong
JY
Han
D000818:Animals; D030801:Animals, Genetically Modified; D015153:Blotting, Western; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens / Q000235:genetics*; D004251:DNA Transposable Elements; D005822:Genetic Vectors; D005854:Germ Cells; D049452:Green Fluorescent Proteins / Q000235:genetics; D019076:Transgenes
D004251:DNA Transposable Elements; D049452:Green Fluorescent Proteins