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Nucleic Acids Research Advance Access originally published online on February 17, 2009
Nucleic Acids Research 2009 37(6):e44; doi:10.1093/nar/gkp058
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Nucleic Acids Research, 2009, Vol. 37, No. 6 e44
© 2009 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

Manipulating transgenes using a chromosome vector

Masashi Ikeno1,2,*, Nobutaka Suzuki2, Yoshinori Hasegawa2 and Tsuneko Okazaki2

1School of Medicine, Keio University, Shinanomachi, Shinjuku-ku, Tokyo 160-8582 and 2Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan

*To whom correspondence should be addressed. Tel: +81 52 385 4259; Fax: +81 52 385 4288; Email: mikeno{at}fujita-hu.ac.jp

Received November 18, 2008. Revised January 19, 2009. Accepted January 20, 2009.

Recent technological advances have enabled us to visualize the organization and dynamics of local chromatin structures; however, the comprehensive mechanisms by which chromatin organization modulates gene regulation are poorly understood. We designed a human artificial chromosome vector that allowed manipulation of transgenes using a method for delivering chromatin architectures into different cell lines from human to fish. This methodology enabled analysis of de novo construction, epigenetic maintenance and changes in the chromatin architecture of specific genes. Expressive and repressive architectures of human STAT3 were established from naked DNA in mouse embryonic stem cells and CHO cells, respectively. Delivery of STAT3 within repressive architecture to embryonic stem cells resulted in STAT3 activation, accompanied by changes in DNA methylation. This technology for manipulating a single gene with a specific chromatin architecture could be utilized in applied biology, including stem cell science and regeneration medicine.


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