Fast and Efficient Expression of Multiple Proteins in Avian Embryos Using mRNA Electroporation

J Vis Exp. 2019 Jun 7:(148). doi: 10.3791/59664.

Abstract

We report that mRNA electroporation permits fluorescent proteins to label cells in living quail embryos more quickly and broadly than DNA electroporation. The high transfection efficiency permits at least 4 distinct mRNAs to be co-transfected with ~87% efficiency. Most of the electroporated mRNAs are degraded during the first 2 h post-electroporation, permitting time-sensitive experiments to be carried out in the developing embryo. Finally, we describe how to dynamically image live embryos electroporated with mRNAs that encode various subcellular targeted fluorescent proteins.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Electroporation / methods*
  • Embryo, Nonmammalian / metabolism*
  • Gene Expression
  • Luminescent Proteins / genetics*
  • Quail / embryology*
  • RNA, Messenger / genetics
  • Time Factors
  • Transfection

Substances

  • Luminescent Proteins
  • RNA, Messenger