Germline transgenesis of the chordate Ciona intestinalis with hyperactive variants of sleeping beauty transposable element.
BACKGROUND
Transposon-mediated transgenesis is an excellent method for creating stable transgenic lines and insertional mutants. In the chordate Ciona intestinalis, Minos is the only transposon that has been used as the tool for germline transformation. Adding another transposon system in this organism enables us to conduct genetic techniques which can only be realized with the use of two transposons.


RESULTS
In the present study, we found that another Tc1/mariner superfamily transposon, sleeping beauty (SB), retains sufficient activity for germline transformation of C. intestinalis. SB shows efficiencies of germline transformation, insertion into gene coding regions, and enhancer detection comparable to those of Minos. We have developed a system for the remobilization of SB copies in the C. intestinalis genome by using transgenic lines expressing SB transposase in the germ cells. With this system, we examined the manner of SB mobilization in the C. intestinalis genome. SB shows intrachromosomal transposition more frequently than Minos.


CONCLUSIONS
SB-based germline transformation and the establishment of a new method that uses its frequent intrachromosomal transposition will result in breakthroughs in genetic approaches that use C. intestinalis together with Minos.
Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka, Japan.
Akiko
A
Hozumi
Kaoru
K
Mita
Csaba
C
Miskey
Lajos
L
Mates
Zsuzsanna
Z
Izsvak
Zoltan
Z
Ivics
Honoo
H
Satake
Yasunori
Y
Sasakura
D000818:Animals; D030801:Animals, Genetically Modified; D002938:Ciona intestinalis / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics*; D018274:Electroporation; D018014:Gene Transfer Techniques; D005854:Germ Cells / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008845:Microinjections; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019895:Transposases / Q000378:metabolism
D017931:DNA Primers; D004251:DNA Transposable Elements; D049452:Green Fluorescent Proteins; D019895:Transposases; C494471:sleeping beauty transposase, human