Germline transgenesis of the ascidian Ciona intestinalis by electroporation.
Microinjection of the Minos transposon is the only reported technique for generating stable transgenic lines in the cosmopolitan ascidian, Ciona intestinalis. To establish a more amenable method for generating stable transgenic Ciona, we examined the possibility of using electroporation of DNA into eggs. From 0-44.4% of electroporated individuals transmitted transgenes to the next generation. The transgene was integrated into one chromosome and multiple copies of the transgene were inserted into one site of the chromosome, indicating that electroporation is an easy and powerful technique for achieving stable transgenesis in C. intestinalis. Together with possible inland culture of this ascidian, this technique will be useful for generating stable lines which have reporter gene expression in a specific tissue or organ and the generation of transposase-expressing stable transgenic (jump-starter) lines and mutator lines which contain a lot of Minos transposons in an insertion position.
Matsuoka
Terumi
T
Department of Zoology, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto, Japan.
Awazu
Satoko
S
Shoguchi
Eiichi
E
Satoh
Nori
N
Sasakura
Yasunori
Y
D000818:Animals; D030801:Animals, Genetically Modified; D002938:Ciona intestinalis / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics; D018274:Electroporation / Q000379:methods*; D005260:Female; D005821:Genetic Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D011993:Recombinant Fusion Proteins / Q000235:genetics
D004251:DNA Transposable Elements; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins