Ubiquitous GFP expression in transgenic chickens using a lentiviral vector.
We report the first ubiquitous green fluorescent protein expression in chicks using a lentiviral vector approach, with eGFP under the control of the phosphoglycerol kinase promoter. Several demonstrations of germline transmission in chicks have been reported previously, using markers that produce tissue-specific, but not ubiquitous, expression. Using embryos sired by a heterozygous male, we demonstrate germline transmission in the embryonic tissue that expresses eGFP uniformly, and that can be used in tissue transplants and processed by in situ hybridization and immunocytochemistry. Transgenic tissue is identifiable by both fluorescence microscopy and immunolabeling, resulting in a permanent marker identifying transgenic cells following processing of the tissue. Stable integration of the transgene has allowed breeding of homozygous males and females that will be used to produce transgenic embryos in 100% of eggs laid upon reaching sexual maturity. These results demonstrate that a transgenic approach in the chick model system is viable and useful even though a relatively long generation time is required. The transgenic chick model will benefit studies on embryonic development, as well as providing the pharmaceutical industry with an economical bioreactor.
Chapman
Susan C
SC
University of Utah School of Medicine, Department of Neurobiology and Anatomy, and Children's Health Research Center, Room 401 MREB, 20 North 1900 East Salt Lake City, Utah 84132-3401, USA. susan.chapman@utah.edu
Lawson
Aaron
A
Macarthur
William C
WC
Wiese
Russell J
RJ
Loechel
Robert H
RH
Burgos-Trinidad
Maria
M
Wakefield
John K
JK
Ramabhadran
Ram
R
Mauch
Teri Jo
TJ
Schoenwolf
Gary C
GC
D000818:Animals; D030801:Animals, Genetically Modified; D002642:Chick Embryo; D002645:Chickens; D047108:Embryonic Development; D005260:Female; D005819:Genetic Markers; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006579:Heterozygote; D006720:Homozygote; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D008856:Microscopy, Fluorescence; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D014018:Tissue Distribution; D019076:Transgenes
D005819:Genetic Markers; D049452:Green Fluorescent Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); C023916:phosphoglycerol kinase