Retroviral Vector-Mediated Gene Transfer into the Chick Optic Vesicle by In Ovo Electroporation

  • Hiraki Sakuta
  • Ryoko Suzuki
  • Masaharu Noda

The chick embryo offers many advantages for developmental studies over other vertebrate embryos as it allows easy access for in ovo surgical manipulations, such as tissue transplantation and the implantation of cultured cells or chemically treated beads for the local release of humoral factors. In particular, owing to its external position in the embryo, the chick eye is a popular model for studying the patterning mechanism of the central nervous system (CNS). This patterning has a crucial role in shaping functional organization because it is the basis of the specific wiring in the CNS. Genetic analysis is not easy in the chick, as compared with the mouse for which transgene introduction or gene targeting techniques have been well established. However, because methods for the expression of exogenous genes and for gene silencing in the chick embryo have been recently developed, the functional analysis of genes has become possible in combination with classical techniques of developmental biology and neurobiology.

One general method is replication-competent retroviral particle-mediated gene transfer in the chick embryo (Morgan and Fekete, 1996). After the infection of viral particles, the viral RNA is reverse-transcribed and integrated into the host genome. Moreover, because the replication-competent viral vectors harbor all of the genome sequences essential for the production of infectious viral particles, horizontal infection continues to the neighboring cells in the host through reproduction of viral particles. Thus, the stable and efficient expression of a transgene can be achieved by this method. However, the system has several restrictions. For example, only coding sequences of less than 2.5 kb can be packaged into the vector. Also, it takes over 24 h for the proteins encoded in the retroviral vectors to be adequately expressed after the injection of viral particles. Therefore, this method is not suitable for the functional analysis of genes in the early developmental stages.


Enhanced Green Fluorescence Protein Long Terminal Repeat Chick Embryo Enhanced Green Fluorescence Protein Expression Optic Vesicle 
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Copyright information

© Springer 2009

Authors and Affiliations

  1. 1.Division of Molecular Neurobiology, National Institute for Basic Biology, and School of Life ScienceThe Graduate University for Advanced StudiesMyodaiji-choJapan

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