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Nat Protoc. 2008;3(3):419-26. doi: 10.1038/nprot.2008.10.

Spatially and temporally controlled electroporation of early chick embryos.

Author information

1
Department of Anatomy and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK. o.voiculescu@ucl.ac.uk

Abstract

The introduction of in ovo electroporation a decade ago has helped the chick embryo to become a powerful system to study gene regulation and function during development. Although this is a simple procedure for embryos of 2-d incubation, earlier stages (from laying to early neurulation, 0-1 d) present special challenges. Here we describe a robust and reproducible protocol for electroporation of expression vectors and morpholino oligonucleotides into the epiblast of embryos from soon after laying (stage XI) to stages 6-7 (early neurulation), with precise spatial and temporal control. Within 3 h, about 12 embryos can be electroporated and set up for culture by the New technique; the effects of morpholinos can be assessed immediately after electroporation, and robust overexpression from plasmid DNA is seen 2-3 h after electroporation. These techniques can be used for time-lapse imaging, gain- and loss-of-function experiments and studying gene regulatory elements in living embryos.

PMID:
18323813
DOI:
10.1038/nprot.2008.10
[Indexed for MEDLINE]

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