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First published online 5 December 2007
doi: 10.1242/dev.008490

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Crucial role of antisense transcription across the Xist promoter in Tsix-mediated Xist chromatin modification

Tatsuya Ohhata^1,2,*, Yuko Hoki^1,2, Hiroyuki Sasaki^2,3 and Takashi Sado^1,2,3,

¹ PRESTO, Japan Science and Technology Agency (JST), 4-1-8, Honcho, Kawaguchi, Saitama, 332-0012, Japan.
² Division of Human Genetics, National Institute of Genetics, Research Organization of Information and Systems, 1111 Yata, Mishima, 411-8540, Japan.
³ Department of Genetics, the Graduate University for Advanced Studies, 1111, Yata, Mishima, 411-8540, Japan.

Author for correspondence (e-mail: tsado{at}lab.nig.ac.jp)

Accepted 12 October 2007

Expression of Xist, which triggers X inactivation, is negatively regulated in cis by an antisense gene, Tsix, transcribed along the entire Xist gene. We recently demonstrated that Tsix silences Xist through modification of the chromatin structure in the Xist promoter region. This finding prompted us to investigate the role of antisense transcription across the Xist promoter in Tsix-mediated silencing. Here, we prematurely terminated Tsix transcription before the Xist promoter and addressed its effect on Xist silencing in mouse embryos. We found that although 93% of the region encoding Tsix was transcribed, truncation of Tsix abolished the antisense regulation of Xist. This resulted in a failure to establish the repressive chromatin configuration at the Xist promoter on the mutated X, including DNA methylation and repressive histone modifications, especially in extraembryonic tissues. These results suggest a crucial role for antisense transcription across the Xist promoter in Xist silencing.

Key words: X inactivation, Antisense regulation, Chromatin modification, Gene targeting, Mouse embryos

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