High-level expression of single-chain Fv-Fc fusion protein in serum and egg white of genetically manipulated chickens by using a retroviral vector.
We report here the generation of transgenic chickens using a retroviral vector for the production of recombinant proteins. It was found that the transgene expression was suppressed when a Moloney murine leukemia virus-based retroviral vector was injected into chicken embryos at the blastodermal stage. When a concentrated viral solution was injected into the heart of developing embryos after 50 to 60 h of incubation, transgene expression was observed throughout the embryo, including the gonads. For practical production, a retroviral vector encoding an expression cassette of antiprion single-chain Fv fused with the Fc region of human immunoglobulin G1 (scFv-Fc) was injected into chicken embryos. The birds that hatched stably produced scFv-Fc in their serum and eggs at high levels (approximately 5.6 mg/ml). We obtained transgenic progeny from a transgenic chicken generated with this procedure. The transgene was stably integrated into the chromosomes of transgenic progeny. The transgenic progeny also expressed scFv-Fc in the serum and eggs.
Kamihira
Masamichi
M
Department of Biotechnology, Graduate School of Engineering, Nagoya University, Chikusa-ku, Japan.
Ono
Ken-ichiro
K
Esaka
Kazuhisa
K
Nishijima
Ken-ichi
K
Kigaku
Ryoko
R
Komatsu
Hiroyuki
H
Yamashita
Takashi
T
Kyogoku
Kenji
K
Iijima
Shinji
S
D000818:Animals; D030801:Animals, Genetically Modified; D000906:Antibodies / Q000097:blood / Q000378:metabolism*; D001756:Blastoderm; D002642:Chick Embryo; D002645:Chickens / Q000235:genetics* / Q000378:metabolism; D004529:Egg White / Q000032:analysis; D005260:Female; D005822:Genetic Vectors; D006321:Heart; D007141:Immunoglobulin Fc Fragments / Q000032:analysis / Q000378:metabolism*; D007128:Immunoglobulin Fragments / Q000032:analysis / Q000378:metabolism*; D008222:Lymphokines / Q000032:analysis / Q000378:metabolism*; D008297:Male; D008979:Moloney murine leukemia virus / Q000235:genetics*; D010063:Ovum / Q000378:metabolism*; D011328:Prions / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D012190:Retroviridae / Q000235:genetics*; D012795:Sialoglycoproteins / Q000032:analysis / Q000378:metabolism*
D000906:Antibodies; C089422:Fv protein, human; D007141:Immunoglobulin Fc Fragments; D007128:Immunoglobulin Fragments; D008222:Lymphokines; D011328:Prions; D011994:Recombinant Proteins; D012795:Sialoglycoproteins