avian adeno-associated virus vector efficiently transduces neurons in the embryonic and post-embryonic chicken brain禽腺相关病毒载体高效能传感神经细胞在胚胎和post-embryonic鸡的大脑.pdfVIP

Avian Adeno-Associated Virus Vector Efficiently Transduces Neurons in the Embryonic and Post- Embryonic Chicken Brain 1 3 1,2 Ryosuke Matsui , Yasuto Tanabe , Dai Watanabe * 1 Department of Molecular and Systems Biology, Graduate School of Biostudies, Kyoto University, Kyoto, Japan, 2 Department of Biological Sciences, Faculty of Medicine, Kyoto University, Kyoto, Japan, 3 Department of Developmental Neuroscience, Graduate School of Frontier Biosciences, Osaka University, Suita, Japan Abstract The domestic chicken is an attractive model system to explore the development and function of brain circuits. Electroporation-mediated and retrovirus (including lentivirus) vector-mediated gene transfer techniques have been widely used to introduce genetic material into chicken cells. However, it is still challenging to efficiently transduce chicken postmitotic neurons without harming the cells. To overcome this problem, we searched for a virus vector suitable for gene transfer into chicken neurons, and report here a novel recombinant virus vector derived from avian adeno-associated virus (A3V). A3V vector efficiently transduces neuronal cells, but not non-neuronal cells in the brain. A single A3V injection into a postembryonic chick brain allows gene expression selectively in neuronal cells within 24 hrs. Such rapid and neuron-specific gene transduction raises the possibility that A3V vector can be utilized for studies of memory formation in filial imprinting, which occurs during the early postnatal days. A3V injection into the neural tube near the ear vesicle at early embryonic stage resulted in persistent and robust gene expression until E20.5 in the auditory brainstem. We further devised an A3V- mediated tetracycline (Tet) depende