Chapman Susan C, Lawson Aaron, Macarthur William C, Wiese Russell J, Loechel Robert H, Burgos-Trinidad Maria, Wakefield John K, Ramabhadran Ram, Mauch Teri Jo, Schoenwolf Gary C
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.
Development. 2005 Mar;132(5):935-40. doi: 10.1242/dev.01652. Epub 2005 Jan 26.
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.
我们报道了首次使用慢病毒载体方法在雏鸡中实现普遍的绿色荧光蛋白表达,其中增强型绿色荧光蛋白(eGFP)受磷酸甘油激酶启动子的控制。此前已有多项使用产生组织特异性而非普遍表达标记的研究报道了雏鸡中的种系传递。利用杂合雄性所产的胚胎,我们在均匀表达eGFP的胚胎组织中证明了种系传递,该组织可用于组织移植,并可通过原位杂交和免疫细胞化学进行处理。转基因组织可通过荧光显微镜和免疫标记进行识别,从而在组织处理后形成识别转基因细胞的永久性标记。转基因的稳定整合使得能够培育出纯合雄性和雌性,它们在达到性成熟后所产的100%的卵中将用于产生转基因胚胎。这些结果表明,尽管需要相对较长的世代时间,但在雏鸡模型系统中采用转基因方法是可行且有用的。转基因雏鸡模型将有益于胚胎发育研究,同时为制药行业提供一种经济的生物反应器。