Improved mRNA electroporation method for Xenopus neurula embryos

✍ Scribed by Satoru Sasagawa; Takashi Takabatake; Yuka Takabatake; Tatsuo Muramatsu; Kazuhito Takeshima

• Book ID: 102843527
• Publisher: John Wiley and Sons
• Year: 2002
• Tongue: English
• Weight: 591 KB
• Volume: 33
• Category: Article
• ISSN: 1526-954X
• DOI: 10.1002/gene.10094

✦ Synopsis

Electroporation has led to new approaches to the analysis of gene regulation of the chick embryonic system. However, application of this method to Xenopus, another model organism of embryology, has left many difficulties to be overcome. The specially devised electrodes, the examination of luciferase activities expressed, and the direct visualization of green fluorescence protein allow us to optimize the conditions of electroporation for Xenopusembryos. The use of mRNA rather than DNA improved the expression efficiency 120 times more than for the case of plasmid DNA, and the effect emerged more immediately after electroporation. The noncontact electroporation adopted here caused less damage to cells and tissues than with the needle type electrode, making it practical for efficient application to early embryos. Furthermore, the mRNA electroporation technique is applicable for other systems in which the DNA electroporation has not had any significant effect because of its low expression efficiency.