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Nucleic Acids Research Advance Access published online on September 15, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp686
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© The Author(s) 2009. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

PiggyBac transgenic strategies in the developing chicken spinal cord

Yanyan Lu, Chengyi Lin and Xiaozhong Wang*

Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, IL60208, USA

*To whom correspondence should be addressed. Tel: +1 847 467 4897; Fax: +1 847 467 1380; Email: awang{at}northwestern.edu

Received April 29, 2009. Revised August 3, 2009. Accepted August 4, 2009.

The chicken spinal cord is an excellent model for the study of early neural development in vertebrates. However, the lack of robust, stable and versatile transgenic methods has limited the usefulness of chick embryos for the study of later neurodevelopmental events. Here we describe a new transgenic approach utilizing the PiggyBac (PB) transposon to facilitate analysis of late-stage neural development such as axon targeting and synaptic connection in the chicken embryo. Using PB transgenic approaches we achieved temporal and spatial regulation of transgene expression and performed stable RNA interference (RNAi). With these new capabilities, we mapped axon projection patterns of V2b subset of spinal interneurons and visualized maturation of the neuromuscular junction (NMJ). Furthermore, PB-mediated RNAi in the chick recapitulated the phenotype of loss of agrin function in the mouse NMJ. The simplicity and versatility of PB-mediated transgenic strategies hold great promise for large-scale genetic analysis of neuronal connectivity in the chick.


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