High-throughput enhancer trap by remobilization of transposon Minos in Ciona intestinalis.
The enhancer trap approach utilizing transposons yields us information about gene functions and gene expression patterns. In the ascidian Ciona intestinalis, transposon-based transgenesis and insertional mutagenesis were achieved with a Tc1/mariner transposon Minos. We report development of a novel technique for enhancer trap in C. intestinalis. This technique uses remobilization of Minos in the Ciona genome. A Minos vector for enhancer trap was constructed and a tandem array insertion of the vector was introduced into the Ciona genome to create a mutator line. Minos was remobilized in Ciona chromosomes to create new insertions by providing transposases. These transposase-introduced animals were crossed with wild-type animals. Nearly 80% of F1 families showed novel GFP expression patterns. This high-throughput enhancer trap screen will be useful to create new marker transgenic lines showing reporter gene expression in specific tissues and to identify novel patterns of gene expression.
Awazu
Satoko
S
Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka, Japan. satoko.awazu@sars.uib.no
Matsuoka
Terumi
T
Inaba
Kazuo
K
Satoh
Nori
N
Sasakura
Yasunori
Y
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D002938:Ciona intestinalis / Q000235:genetics*; D004247:DNA / Q000235:genetics; D004251:DNA Transposable Elements; D004742:Enhancer Elements, Genetic; D015870:Gene Expression; D017930:Genes, Reporter; D005821:Genetic Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence; D011994:Recombinant Proteins / Q000235:genetics; D019895:Transposases / Q000235:genetics
D004251:DNA Transposable Elements; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; C473499:Minos transposase; D019895:Transposases