High-throughput enhancer trap by remobilization of transposon Minos in Ciona intestinalis

Authors

  • Satoko Awazu,

    Corresponding author
    1. Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka, Japan
    2. Department of Zoology, Graduate School of Science, Kyoto University, Sakyo-Ku, Kyoto, Japan
    Current affiliation:
    1. Sars International Centre for Marine Molecular Biology, University of Bergen, Thormøhlensgate 55, 5008 Bergen, Norway
    • Sars International Centre for Marine Molecular Biology, University of Bergen, Thormøhlensgate 55, 5008 Bergen, Norway
    Search for more papers by this author
  • Terumi Matsuoka,

    1. Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka, Japan
    2. Department of Zoology, Graduate School of Science, Kyoto University, Sakyo-Ku, Kyoto, Japan
    Search for more papers by this author
  • Kazuo Inaba,

    1. Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka, Japan
    Search for more papers by this author
  • Nori Satoh,

    1. Department of Zoology, Graduate School of Science, Kyoto University, Sakyo-Ku, Kyoto, Japan
    2. CREST, Japan Science and Technology Agency, Kawaguchi, Saitama, Japan
    Search for more papers by this author
  • Yasunori Sasakura

    1. Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka, Japan
    Search for more papers by this author

Abstract

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. genesis 45:307–317, 2007. © 2007 Wiley-Liss, Inc.

Ancillary