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piggyBac transposition into primordial germ cells is an efficient tool for transgenesis in chickens

Author:
Park, Tae Sub, Han, Jae Yong
Source:
Proceedings of the National Academy of Sciences of the United States of America 2012 v.109 no.24 pp. 9337-9341
ISSN:
0027-8424
Subject:
biotechnology, chickens, chicks, chromosomes, gene expression, germ cells, heterozygosity, medicine, poultry production, progeny, quails, transgenes, transgenic animals, transposition (genetics)
Abstract:
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 piggyBa c transposition into the chicken PGC line could be the surest way to generate transgenic chickens safely for practical applications.
Agid:
1721190