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Published Online September 16, 2010
% B/ `4 N8 ^4 g/ BScience DOI: 10.1126/science.1194174
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Impeding Xist Expression from the Active X Chromosome Improves Mouse Somatic Cell Nuclear Transfer
9 I) B$ o  r% o0 xKimiko Inoue,1,2 Takashi Kohda,3 Michihiko Sugimoto,1 Takashi Sado,4 Narumi Ogonuki,1 Shogo Matoba,1 Hirosuke Shiura,1 Rieko Ikeda,1 Keiji Mochida,1 Takashi Fujii,5 Ken Sawai,5 Arie P. Otte,6 X. Cindy Tian,7 Xiangzhong Yang,7 Fumitoshi Ishino,3 Kuniya Abe,1,2 Atsuo Ogura1,2,8,* . I6 a9 g& r* [* K+ N" ?) {) @
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Cloning mammals by somatic cell nuclear transfer (SCNT) is highly inefficient because of erroneous reprogramming of the donor genome. Reprogramming errors appear to arise randomly, but the nature of nonrandom, SCNT-specific errors remains elusive. We found that Xist, a noncoding RNA that inactivates one of the two X chromosomes in females, was ectopically expressed from the active X (Xa) chromosome in cloned mouse embryos of both sexes. Deletion of Xist on Xa showed in normal global gene expression and resulted in about an 8- to 9-fold increase in cloning efficiency. We also identified an Xist-independent mechanism that specifically downregulated a subset of X-linked genes through somatic-type repressive histone blocks. Thus, we have identified nonrandom reprogramming errors in mouse cloning that can be altered to improve the efficiency of SCNT methods.
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1 BioResource Center, RIKEN, Tsukuba, Japan.% M& x8 V- V$ h* ]% _8 a
2 Graduate School of Life and Environmental Science, University of Tsukuba, Tsukuba, Japan.9 y5 R3 p+ E- z+ ?6 y
3 Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan.0 U8 _/ i1 C# F: H
4 Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
" l6 p+ J# p; ~9 s! W$ Y8 G, L3 E5 Faculty of Agriculture, Iwate University, Iwate, Japan.
/ N) q3 R  K# M: t6 Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, Netherlands.
0 F! F5 B, k$ D0 J" ]3 H" U! C7 Center for Regenerative Biology and Department of Animal Science, University of Connecticut, Connecticut 06269, USA.: S& X4 t1 r( M& @: g+ f
8 The Center for Disease Biology and Integrative Medicine, Faculty of Medicine, University of Tokyo, Tokyo, Japan. - D8 o: p. H7 \9 b' x: a
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* To whom correspondence should be addressed. E-mail: ogura@rtc.riken.go.jp

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