Spatially and Temporally Controlled Electroporation of Early Chick Embryos

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chick Embryo
  • Electroporation / methods*
  • Embryo Culture Techniques
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation, Enzymologic / genetics
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • Oligonucleotides, Antisense / genetics
  • Oligonucleotides, Antisense / metabolism
  • Time Factors

Substances

  • Oligonucleotides, Antisense