Efficient gene transfer by electroporation of chick embryos in ovo has allowed the development of new approaches to the analysis of gene regulation, function and expression, creating an exciting opportunity to build upon the classical manipulative advantages of the chick embryonic system. This method is applicable to other vertebrate embryos and is an important tool with which to address cell and developmental biology questions. Here we describe the technical aspects of in ovo electroporation, its different applications and future perspectives.
This is a preview of subscription content, access via your institution
Open Access articles citing this article.
Cellular and Molecular Life Sciences Open Access 05 November 2020
Scientific Reports Open Access 24 July 2019
Neural Development Open Access 08 March 2018
Subscribe to Journal
Get full journal access for 1 year
only $8.25 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Le Douarin, N. A biological cell labeling technique and its used in experimental embryology. Dev. Biol. 30, 217–222 (1973).
Couly, G. F., Coltey, P. M. & Le Douarin, N. M. The developmental fate of the cephalic mesoderm in quail-chick chimeras. Development 114, 1–15 (1992).
Couly, G. F., Coltey, P. M. & Le Douarin, N. M. The triple origin of skull in higher vertebrates — a study in quail-chick chimeras. Development 117, 409–429 (1993).
Bronner-Fraser, M. & Fraser, S. Cell lineage analysis reveals multipotency of some avian neural crest cells. Nature 335, 161–164 (1988).
Fraser, S., Keynes, R. & Lumsden, A. Segmentation in the chick embryo hindbrain is defined by cell lineage restrictions. Nature 344, 431–435 (1990).
Krull, C. E., Collazo, A., Fraser, S. E. & Bronner-Fraser, M. Segmental migration of trunk neural crest: time lapse analysis reveals a role for PNA-binding molecules. Development 121, 3733–3743 (1995).
Kulesa, P. Neural crest cell dynamics revealed by time-lapse video microscopy of whole chick explant cultures. Dev. Biol. 204, 327–344 (1998).
Petropoulos, C. & Hughes, S. Replication-competent retrovirus vectors for the transfer and expression of gene cassettes in avian cells. J. Virol. 65, 3728– 3737 (1991).
Petropoulos, C., Payne, W., Salter, D. & Hughes, S. Using avian retroviral vectors for gene transfer. J. Virol. 66, 3391–3397 (1992).
Shillito, R., Saul, M., Paszkowski, J., Muller, M. & Potrykus, I. High efficiency direct gene transfer to plants. Bio/Technol. 3, 1099–1103 (1985).
Andreason, G. & Evans, G. Induction and expression of DNA molecules in eukaryotic cells by electroporation. Biotechniques 6, 650–660 (1988).
Takahashi, M. et al. Gene transfer into human leukemia cell lines by electroporation: experience with exponentially decaying and square wave pulse. Leukemia Res. 15, 507–513 (1991).
Muramatsu, T., Mizutani, Y., Ohmori, Y. & Okumura, J.-i. Comparison of three non-viral transfection methods for foreign gene expression in early chicken embryos in ovo. Biochem. Biophys. Res. Comum. 230, 376–380 (1997).
Nishi, T. et al. High-efficiency in vivo gene transfer using intraarterial plasmid DNA injection following in vivo electroporation. Cancer Res. 56, 1050–1055 (1996).
Momose, T. et al. Efficient targeting of gene expression in chick embryos by microelectroporation. Dev. Growth Differ. 41, 335–344 (1999).
Suemori, H. et al. A mouse embryonic stem cell line showing pluripotency of differentiation in early embryos and ubiquitous β-galactosidase expression. Cell Differ. Dev. 29, 181–186 (1990).
Gould, A., Itasaki, N. & Krumlauf, R. Initiation of rhombomeric Hoxb4 expression requires induction by somites and a retinoid pathway. Neuron 21, 39–51 (1998).
Funahashi, J.-i. et al. Role of Pax5 in the regulation of a mid-hindbrain organizer’s activity. Dev. Growth Differ. 41, 59– 72 (1999).
Akamatsu, W. et al. Mammalian ELAV-like neuronal RNA-binding proteins HuB and HuC promote neuronal development in both the central and the peripheral nervous systems. Proc. Natl Acad. Sci. USA 96, 9885–9890 (1999).
Ogino, H. & Yasuda, K. Induction of lens differentiation by activation of a bZIP transcription factor, L-Maf. Science 280, 115–118 (1998).
Takeuchi, J. K. et al. Tbx5 and Tbx4 genes determine the wing/leg identity of limb buds. Nature 398, 810– 814 (1999).
Manzanares, M. et al. Conserved and distinct roles of kreisler in regulation of paralogous Hoxa3 and Hoxb3 genes. Development 126, 759–769 (1999).
Morrison, A. et al. Comparative analysis of Hoxb-4 regulation in transgenic mice. Mech. Dev. 53, 47– 59 (1995).
Morrison, A., Ariza-McNaughton, L., Gould, A., Featherstone, M. & Krumlauf, R. HOXD4 and regulation of the group 4 paralog genes. Development 124, 3135–3146 (1997).
Whiting, J. et al. Multiple spatially specific enhancers are required to reconstruct the pattern of Hox-2.6 gene expression. Genes Dev. 5, 2048–2059 (1991).
Aparicio, S. et al. Detecting conserved regulatory elements with the model genome of the Japanese puffer fish Fugu rubripes. Proc. Natl Acad. Sci. USA 92, 1684–1688 (1995).
Pöpperl, H. . et al. Segmental expression of Hoxb1 is controlled by a highly conserved autoregulatory loop dependent upon exd/Pbx. Cell 81, 1031–1042 (1995).
Studer, M., Pöpperl, H., Marshall, H., Kuroiwa, A. & Krumlauf, R. Role of a conserved retinoic acid response element in rhombomere restriction of Hoxb-1. Science 265, 1728–1732 (1994).
Aihara, H. & Miyazaki, J.-i. Gene transfer into muscle by electroporation in vivo. Nature Biotech. 16, 867–870 (1998).
Rols, M.-P. et al. In vivo electrically mediated protein and gene transfer in murine melanoma. Nature Biotech. 16, 168–171 (1998).
Sundin, O. & Eichele, G. A homeo domain protein reveals the metameric nature of the developing chick hindbrain. Genes Dev. 4, 1267–1276 (1990).
Maden, M. et al. Retinoic acid-binding protein and homeobox expression in rhombomeres of the chick embryo. Development 111, 35–44 (1991).
We thank M. Manzanares for constructing the Hoxa3 vectors and help in testing their activity; A. Morrison, S. Nonchev, H. Popperl, M. Studer and H. Marshall for the region-specific enhancer constructs; and K. Kusumi, T. Jinks and M. Martínez-Pastor for discussions and testing approaches for electroporation. N.I. thanks H. Nakamura, J.-i. Funahashi and N. Osumi for technical suggestions; and Y. Imada and Y. Hayakawa for technical support. S.B.-V. was supported by fellowships from the French Cancer Research Association (ASC) and EMBO; N.I. was supported by an HFSP fellowship and the MRC; R.K.’s research was funded by the MRC.
About this article
Cite this article
Itasaki, N., Bel-Vialar, S. & Krumlauf, R. ‘Shocking’ developments in chick embryology: electroporation and in ovo gene expression . Nat Cell Biol 1, E203–E207 (1999). https://doi.org/10.1038/70231
This article is cited by
Cellular and Molecular Life Sciences (2021)
Science China Life Sciences (2021)
Scientific Reports (2019)
Neural Development (2018)
Cellular and Molecular Neurobiology (2018)