Protocol abstract
Nature Protocols 3, - 419 - 426 (2008)
Published online: 21 February 2008 | Corrected online: 4 September 2008 | doi:10.1038/nprot.2008.10
Subject Categories: Cell and developmental biology | Cell and tissue culture | Genetic modification | Model organisms
Spatially and temporally controlled electroporation of early chick embryos
Octavian Voiculescu1, Costis Papanayotou1 & Claudio D Stern1
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.
- Department of Anatomy and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK.
Correspondence to: Octavian Voiculescu1 e-mail: o.voiculescu@ucl.ac.uk
Correspondence to: Claudio D Stern1 e-mail: c.stern@ucl.ac.uk
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