Predictable mosaic transgene expression in ascidian embryos produced with a simple electroporation device.
Two customized electroporators were specifically designed for creating transgenic ascidian embryos. These electroporators were simple to build, inexpensive, and produced transgenic embryos with efficiencies that equaled or rivaled commercially available machines. A key design feature of these machines resulted in the generation of consistent electroporation pulses providing repeatability between experiments. These devices were used to optimize experimental parameters allowing for the creation of transient transgenic embryos with predictable patterns of mosaic transgene expression. We used these new electroporators to examine the expression of two different fluorescent protein reporter genes with regard to embryonic cell lineage. In general, transgene expression followed the embryonic cell lineage and coelectroporated transgenes were always expressed in the same embryonic cells. Our analysis also indicated that, during development, transgenes could be lost from embryonic cells, suggesting that transgenes may be present in extrachromosomal arrays, as has been observed in other organisms. Our new electroporator designs will allow ascidian researchers to inexpensively produce transgenic ascidians and should prove useful for adapting the electroporation technique to other marine embryo systems.
Zeller
Robert W
RW
Molecular Biology Institute and Coastal and Marine Institute, San Diego State University, San Diego, California 92182-4614, USA. rzeller@sciences.sdsu.edu
Virata
Michael J
MJ
Cone
Angela C
AC
D000818:Animals; D030801:Animals, Genetically Modified; D019070:Cell Lineage; D018274:Electroporation / Q000295:instrumentation / Q000379:methods*; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism; D017930:Genes, Reporter; D019076:Transgenes; D014561:Urochordata / Q000166:cytology / Q000235:genetics* / Q000378:metabolism