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Journal of Biochemistry 2006 139(4):705-714; doi:10.1093/jb/mvj075
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© 2006 The Japanese Biochemical Society.

Regular Paper

Ars Insulator Identified in Sea Urchin Possesses an Activity to Ensure the Transgene Expression in Mouse Cells

Shoji Tajima1,*, Keiko Shinohara1, Maiko Fukumoto1, Reiko Zaitsu1, Junichi Miyagawa1, Shinjiro Hino2, Jun Fan2, Koji Akasaka3 and Masao Matsuoka2

1 Laboratory of Epigenetics, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871; 2 Laboratory of Virus Immunology, Institute for Virus Research, Kyoto University, 53 Shogoin-Kawahara-cho, Sakyo-ku, Kyoto 606-8507; and 3 Misaki Marine Biological Station, Graduate School of Science, The University of Tokyo, 1024 Koajiro, Miura, Kanagawa 238-0225

* To whom correspondence should be addressed. Tel: +81-6-6879-8627, Fax: +81-6-6879-8629, E-mail: tajima{at}protein.osaka-u.ac.jp

Sea urchin arylsulfatase (Ars) gene locus has features of an insulator, i.e., blocking of enhancer and promoter interaction, and protection of a transgene against positional effects [Akasaka et al. (1999) Cell. Mol. Biol. 45, 555–565]. To examine the effect of Ars insulator on long-term expression of a transgene, the insulator was inserted into LTR of retrovirus vector harboring hrGFP gene as a reporter, and then introduced into mouse myoblast cells. The isolated clones transduced with the reporter gene with or without Ars insulator were cultured for more than 20 wk in the absence of a selection reagent, and the expression of hrGFP was periodically determined. Expression of hrGFP in four clones transduced with the reporter gene without Ars insulator was completely silenced after 20 wk of culture. On the other hand, hrGFP was expressed in all clones with Ars insulator inserted in one of the two different orientations. Histone H3 deacetylation and DNA methylation of the 5'LTR promoter region, signs for heterochromatin and silencing, were suppressed in the clones that were expressing hrGFP. Ars insulator is effective in maintaining a transgene in mouse cells in an orientation-dependent manner, and will be a useful tool to ensure stable expression of a transgene.


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