Article Developmental Biology Combinational electroporation and transplantation approach to studying gene functions in avian embryos Xiaoyu Wang • Yan Li • Guang Wang • Andrea Mu ¨ nsterberg • Manli Chuai • KaHo Kenneth Lee • Lijing Wang • Xuesong Yang Received: 22 April 2013 / Accepted: 19 June 2013 / Published online: 29 January 2014 Ó Science China Press and Springer-Verlag Berlin Heidelberg 2014 Abstract Gene transfection is an indispensable approach for studying gene function since it provides important information on gain- and/or loss-of-function. Chick embryos are also extensively employed for studying bio- logical function since they are easily accessible and can be maintained alive after manipulation. The combination of both techniques presents a powerful approach to under- standing how genes regulate embryo development. Fur- thermore, combining these approaches with tissue transplant techniques make even more attractive for elu- cidate gene function. Electroporation, employing parallelly fashioned electrodes, has been widely used in chick embryos. However, experimenters have been frustrated by unsuccessfully transfection in some embryonic tissue of interest because the electrodes were improperly positioned. We presently demonstrated the different patterns of orga- nizing and positioning the electrodes, in combination with tissue transplantation, to efficiently and specifically trans- fect the chick embryonic head, trunk neural tube, heart tube, somites and neural crest cells with the GFP reporter gene. Keywords Chick embryo Á Electroporation Á Gene transfection Á Transplantation 1 Introduction At present, gain- and loss-of-function studies have been an indispensable approach to understanding how a particular gene functions at multiple levels within an organism [1]. Although numerous gene transfection techniques have been explored, there are only three effective gene transfection techniques that could be applied to chick embryos. These three techniques include lipofection [2–4], virus-mediated transfection [5, 6] and electroporation [7]. The lipofection approach involves delivering liposomes containing the gene of interest by fusion into the cell membrane. How- ever, there have been many reports of unsuccessful attempts to use lipofection to transfect the chick embryo, in spite of its high efficacies in many cell lines. Another disadvantage of the lipofection technique is the lack of control to precisely target rather than mass transfect embryonic tissues. Virus-mediated transfection is based on the uses of viruses to deliver the gene of interest into host cell genome, which could confound the results produced and also require time for the virus to integrate [8]. The electroporation technique delivers DNA plasmid directly into cells by transiently opening small holes in cell mem- brane induced by electrical pulse [9]. Electroporation has X. Wang Á Y. Li Á G. Wang Á KaHo. K. Lee Á X. Yang (&) Department of Histology and Embryology, Key Laboratory for Regenerative Medicine of the Ministry of Education, Medical College of Jinan University, Guangzhou 510632, China e-mail: yang_xuesong@126.com A. Mu ¨nsterberg School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK M. Chuai Division of Cell and Developmental Biology, University of Dundee, Dundee DD1 5EH, UK KaHo. K. Lee Stem Cell and Regeneration Thematic Research Programme, School of Biomedical Sciences, Chinese University of Hong Kong, Shatin, Hong Kong, China L. Wang (&) Institute of Vascular Biological Sciences, Guangdong Pharmaceutical University, Guangzhou 510006, China e-mail: wanglijing62@163.com 123 Chin. Sci. Bull. (2014) 59(7):616–624 csb.scichina.com DOI 10.1007/s11434-013-0090-8 www.springer.com/scp