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This Article Full Text Full Text (PDF) Color Figures 1 and 2 All Versions of this Article: jas.2009-1983v1 87/9/2791    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Google Scholar Articles by Shin, J. Articles by Lee, K. PubMed PubMed Citation Articles by Shin, J. Articles by Lee, K.

Technical note: A gene delivery system in the embryonic cells of avian species using a human adenoviral vector

J. Shin^*,,1, D. R. Bae^*,1, J. D. Latshaw^*, M. P. Wick^*, J. M. Reddish^* and K. Lee^*,,2

^* Department of Animal Sciences, and The Ohio State University Interdisciplinary PhD Program in Nutrition, The Ohio State University, Columbus 43210

² Corresponding author: lee.2626{at}osu.edu

Adenovirus (Ad) has been used in vivo and in vitro as a vector to carry a foreign gene for efficient gene delivery into various cell types and tissues of animals. The aim of the current study was to evaluate the Ad delivery system in primary avian cells. Primary cells isolated from the embryonic pectoralis major muscles of the chicken and quail were cultured and incubated with human recombinant Ad serotype 5 (Ad5) containing sequences encoding either the green fluorescence protein (GFP) gene alone, as a tracking marker, or both GFP and murine 3-hydroxyisobutyryl-CoA hydrolase (mHIBCH) as a target gene. The fluorescent GFP images showed the successful delivery of a target gene using Ad5 in the primary avian cultured cells. In addition, immunostaining of the myosin heavy chain (MyHC) in these cells indicated that a large population of the cells was myogenic. Colocalization of GFP-positive cells with MyHC staining was mostly found in MyHC-negative cells, indicating successful delivery of Ad5 into a large population of mononucleated cells. Furthermore, the current fluorescence study detected the dual expression of GFP and mHIBCH protein in GFP-positive cells. Finally, Western blot analysis confirmed that the Ad-mediated expression of mHIBCH protein was specific in primary cultures of avian myogenic cells and that the mHIBCH protein expression was continued for 15 d after infection in chicken primary cells. These data demonstrate that Ad5 is a feasible tool to express foreign genes in primary cultured cells of avian species, providing a new approach to study the function of genes of interest in muscle development and metabolism.

Key Words: avian primary cell • gene delivery • human recombinant adenovirus