RNAi-mediated Knockdown of <i>Xist</i> Does Not Rescue the Impaired Development of Female Cloned Mouse Embryos
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- OIKAWA Mami
- RIKEN BioResource Center, Ibaraki 305-0074, Japan Department of Epigenetics, Medical Research Institute, Tokyo Medical and Dental University, Tokyo 113-8510, Japan
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- MATOBA Shogo
- RIKEN BioResource Center, Ibaraki 305-0074, Japan
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- INOUE Kimiko
- RIKEN BioResource Center, Ibaraki 305-0074, Japan Graduate School of Life and Environmental Science, University of Tsukuba, Ibaraki 305-8572, Japan
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- KAMIMURA Satoshi
- RIKEN BioResource Center, Ibaraki 305-0074, Japan
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- HIROSE Michiko
- RIKEN BioResource Center, Ibaraki 305-0074, Japan
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- OGONUKI Narumi
- RIKEN BioResource Center, Ibaraki 305-0074, Japan
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- SHIURA Hirosuke
- RIKEN BioResource Center, Ibaraki 305-0074, Japan
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- SUGIMOTO Michihiko
- RIKEN BioResource Center, Ibaraki 305-0074, Japan
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- ABE Kuniya
- RIKEN BioResource Center, Ibaraki 305-0074, Japan
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- ISHINO Fumitoshi
- Department of Epigenetics, Medical Research Institute, Tokyo Medical and Dental University, Tokyo 113-8510, Japan
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- OGURA Atsuo
- RIKEN BioResource Center, Ibaraki 305-0074, Japan Graduate School of Life and Environmental Science, University of Tsukuba, Ibaraki 305-8572, Japan Center for Disease Biology and Integrative Medicine, Faculty of Medicine, University of Tokyo, Tokyo 113-0033, Japan
Bibliographic Information
- Other Title
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- RNAi-mediated Knockdown of Xist Does Not Rescue the Impaired Development of Female Cloned Mouse Embryos
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Abstract
In mice, one of the major epigenetic errors associated with somatic cell nuclear transfer (SCNT) is ectopic expression of Xist during the preimplantation period in both sexes. We found that this aberrant Xist expression could be impeded by deletion of Xist from the putative active X chromosome in donor cells. In male clones, it was also found that prior injection of Xist-specific siRNA could significantly improve the postimplantation development of cloned embryos as a result of a significant repression of Xist at the morula stage. In this study, we examined whether the same knockdown strategy could work as well in female SCNT-derived embryos. Embryos were reconstructed with cumulus cell nuclei and injected with Xist-specific siRNA at 6–7 h after oocyte activation. RNA FISH analysis revealed that siRNA treatment successfully repressed Xist RNA at the morula stage, as shown by the significant decrease in the number of cloud-type Xist signals in the blastomere nuclei. However, blastomeres with different sizes (from “pinpoint” to “cloud”) and numbers of Xist RNA signals remained within single embryos. After implantation, the dysregulated Xist expression was normalized autonomously, as in male clones, to a state of monoallelic expression in both embryonic and extraembryonic tissues. However, at term there was no significant improvement in the survival of the siRNA-injected cloned embryos. Thus, siRNA injection was largely effective in repressing the Xist overexpression in female cloned embryos but failed to rescue them, probably because of an inability to mimic consistent monoallelic Xist expression in these embryos. This could only be achieved in female embryos by applying a gene knockout strategy rather than an siRNA approach.
Journal
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- Journal of Reproduction and Development
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Journal of Reproduction and Development 59 (3), 231-237, 2013
The Society for Reproduction and Development
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Details
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- CRID
- 1390282681314511232
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- NII Article ID
- 10031184751
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- NII Book ID
- AA10936678
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- COI
- 1:STN:280:DC%2BC3szjvVahtQ%3D%3D
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- ISSN
- 13484400
- 09168818
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- NDL BIB ID
- 024616784
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- PubMed
- 23363561
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
- KAKEN
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- Abstract License Flag
- Disallowed