Generation of transgenic quail through germ cell-mediated germline transmission.
Here, we describe the production of transgenic quail via a germline transmission system using postmigratory gonadal primordial germ cells (gPGCs). gPGCs retrieved from the embryonic gonads of 5-day-old birds were transduced with a lentiviral vector and subsequently transferred into recipient embryos. Testcross and genetic analyses revealed that among three germline chimeric G0 quail, one male produced transgenic offspring; of 310 hatchlings from the transgenic germline chimera, 24 were identified as donor-derived offspring, and 6 were transgenic (6/310, 1.9%). Conventional transgenesis using stage X blastodermal embryos was also conducted, but the efficiency of transgenesis was similar between the two systems (<1.6 vs. 1.9% for the conventional and gPGC-mediated systems, respectively). However, substantial advantages can be gained from gPGC-mediated method in that it enables an induced germline modification, whereas direct retroviral transfer to stage X embryos causes mosaic integration. The use of gonadal PGCs for transgenesis may lead to the production of bioreactors.
Shin
Sang Su
SS
Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, Korea.
Kim
Tae Min
TM
Kim
Sun Young
SY
Kim
Tae Wan
TW
Seo
Hee Won
HW
Lee
Seul Ki
SK
Kwon
Se Chang
SC
Lee
Gwan Sun
GS
Kim
Heebal
H
Lim
Jeong Mook
JM
Han
Jae Yong
JY
D000818:Animals; D030801:Animals, Genetically Modified; D001756:Blastoderm / Q000502:physiology; D003370:Coturnix / Q000235:genetics / Q000502:physiology*; D003433:Crosses, Genetic; D004625:Embryo, Nonmammalian / Q000502:physiology; D005241:Feathers / Q000502:physiology; D005260:Female; D005854:Germ Cells / Q000502:physiology; D018095:Germ-Line Mutation; D008297:Male; D010063:Ovum / Q000502:physiology