Generation of transgenic quail through germ cell‐mediated germline transmission

Abstract

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.

The expression profile of apoptosis-related genes in the chicken as a human epithelial ovarian cancer model

“…Through this study, we have provided a new application of the apoptotic status of chicken ovarian cancer as an excellent model of human epithelial ovarian cancer. Through the combination of our transgenic techniques in chickens (35), our findings will be useful in addressing the incidence of cancer due to dysregulated apoptosis. In conclusion, our study provides new insight into the development of therapeutic agents based on the apoptotic pathway in human ovarian cancer.…”

Establishing Germline Chimeric Chickens Using Primordial Germ Cells

“…Thus, it may be possible to produce W chromosome spermtogonia using this method. However, heterosexual (female to male or male to female) transfer of PGCs has been shown to cause abnormal development in the gonads of chimeric chicken embryo (Furuta et al, 1999;2008;Yamaguchi et al, 2000). In avians, estrogen is necessary to initiate feminization in normal females and cytochrome P450 aromatase is an important enzyme for the conversion of testosterone to estrogen.…”

“…PGCs are a useful means for producing transgenic chickens because they can differentiate into gametes. PGCs collected from embryos were cultured in order to transfer exogenous genes and used for the production of transgenic chicks via germinal chimeric chickens (Vick et al, 1993b;Shin et al, 2008;Furuta et al, 2010). Presence and expression of exogenous genes were detected in the next generation (van de Lavoir et al, 2006).…”

The reversible developmental unipotency of germ cells in chicken

“…The other possibility is that PGCs freshly isolated and transferred into adult testis could not overcome the developmental gap for (de)differentiation and maturation or actually, the transplanted PGCs were a mixed population retrieved from male and female embryos which probably affected germ cell differentiation in the recipient testis. Although both the PGC/EGC-to-embryo and SSC/GSC-to-testis germline transmission systems can generate germline chimeras or transgenic animals (van de Lavoir et al 2006, Kalina et al 2007, Shin et al 2008, Han 2009), each possesses different merits. The classical embryo-mediated system is more practical than the testis-mediated system and detailed information is available for the chicken.…”