Spatially and temporally controlled electroporation of early chick embryos |
| |
Authors: | Voiculescu Octavian Papanayotou Costis Stern Claudio D |
| |
Affiliation: | 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. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |
|