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Improved transfection efficiency of chicken gonadal primordial germ cells for the production of transgenic poultry

Cited 26 time in Web of Science Cited 34 time in Scopus
Authors
Hong, Yeong Ho; Kim Moon, Yangha; Jeong, Dong Kee; Han, Jae Yong
Issue Date
1998
Publisher
Springer
Citation
Transgenic Research, vol.7 no.4, pp. 247-252
Keywords
transfectionelectroporationPGCstransgenic poultry
Abstract
Electroporation is a common method of DNA transfection for many types of eukaryotic cells, but has not been
attempted in avian primordial germ cells (PGCs). DNA uptake in chicken primordial germ cells (PGCs) was tested
using electroporation with and without dimethyl sulfoxide (DMSO). Gonadal tissue and chicken embryonic
fibroblasts (CEFs) were isolated from 6-day-old embryos (stage 29), transfected with pCMVâ carrying the bacterial
lacZ gene, and cultured for 24 h. Gonadal primordial germ cells (gPGCs) were purified from culture using a Ficoll
gradient. The addition of DMSO significantly increased the transfection efficiency of gPGCs but had no effect on
chicken embryonic fibroblasts. Electroporation of gPGCs resulted in an 80% transfection efficiency, compared with
about 17% observed with liposomes. Approximately 200 transfected gPGCs were injected into 2.5-day-old (stage
17) recipient embryos and the eggs were incubated for an additional 3.5 days, 7.5 days or to hatching. The
exogenous gene was detectable in 100%, 67% and 41% of the 6-day-old (stage 29), 10-day-old (stage 36) recipient
embryos and hatched chicks gonads, respectively. PCR analysis of DNA from the hatched chicks showed that
exogenous lacZ DNA was detected only in the gonad and not the liver and heart. These results indicated that
electroporation was a suitable means of transfecting avian gPCGs for the goal of producing transgenic poultry.
ISSN
0962-8819
Language
English
URI
https://hdl.handle.net/10371/99050
DOI
https://doi.org/10.1023/A:1008861826681
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College of Agriculture and Life Sciences (농업생명과학대학)Dept. of Food and Animal Biotechnology (식품·동물생명공학부)Journal Papers (저널논문_식품·동물생명공학부)
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