Generation of tissue-specific transgenic birds with lentiviral vectors

Abstract

Birds are of great interest for a variety of research purposes, and effective methods for manipulating the avian genome would greatly accelerate progress in fields that rely on birds as model systems for biological research, such as developmental biology and behavioral neurobiology. Here, we describe a simple and effective method for producing transgenic birds. We used lentiviral vectors to produce transgenic quails that express GFP driven by the human synapsin gene I promoter. Expression of GFP was specific to neurons and consistent across multiple generations. Expression was sufficient to allow visualization of individual axons and dendrites of neurons in vivo by intrinsic GFP fluorescence. Tissuespecific transgene expression at high levels provides a powerful tool for biological research and opens new avenues for genetic manipulation in birds.avian ͉ genetics ͉ lentivirus ͉ synapsin ͉ neuron

Oviduct-specific expression of two therapeutic proteins in transgenic hens

“…Additionally, immunohistochemical staining of sections of the magnum revealed that recombinant protein synthesis was restricted to the region containing the tubular gland cells, the site of ovalbumin synthesis in the magnum, and was absent from adjacent epithelial cells. This observation of tissue-specific transgene expression, using a lentiviral vector to deliver the transgene, is consistent with previous reports from mouse (14), pig (15), quail (9), and rat (16).…”

“…High levels of somatic chimerism with chicken ES cells and germ-line contribution by primordial germ cells were demonstrated, but the fidelity of oviduct-specific expression of a human monoclonal antibody in chicken ES cells chimeras was variable. We have shown that lentiviral vectors can be used to generate germ-line transgenic chickens with high efficiency (8), and tissue-specific expression of a transgene delivered using a lentiviral vector has been demonstrated in quail (9).…”

Evaluation of laser-assisted lentiviral transgenesis in bovine

“…Lentiviral vectors are also suitable to create genetically modified birds. Transgenic chicken stably expressed a reporter gene up to the F2 generation (McGrew et al, 2004), and tissue-specific expression in birds was reported in quails, where Scott et al demonstrated that the use of a GFP vector, driven by the human synapsin gene I promoter lead to the expression of GFP in neurons, which was persistent across multiple generations (Scott and Lois, 2005).…”

Transgenic mice and their impact on kidney research

“…First mice carrying so-called conditional alleles were published in the early 90s [26,27], and, since then, the number of strains carrying conditional alleles or expressing Cre recombinase is steadily increasing. Only recently, another transgenic approach has emerged, using lentiviral vectors [55], that contributed to two major aspects, first allowing to manipulate species other than the mouse [94], and second, to silence gene expression in vivo using siRNA-expressing vectors [80]. Lentivirus-mediated transgenesis can be a very Adding genetic information to the genome: pronuclear injection…”