RNAi-mediated Knockdown of Xist Does Not Rescue the Impaired Development of Female Cloned Mouse Embryos
-
- OIKAWA Mami OIKAWA Mami
- RIKEN BioResource Center
-
- MATOBA Shogo MATOBA Shogo
- RIKEN BioResource Center
-
- INOUE Kimiko [他] INOUE Kimiko
- RIKEN BioResource Center
-
- KAMIMURA Satoshi
- RIKEN BioResource Center
-
- HIROSE Michiko
- RIKEN BioResource Center
-
- OGONUKI Narumi
- RIKEN BioResource Center
-
- SHIURA Hirosuke
- RIKEN BioResource Center
-
- SUGIMOTO Michihiko
- RIKEN BioResource Center
-
- ABE Kuniya
- RIKEN BioResource Center
-
- ISHINO Fumitoshi
- Department of Epigenetics, Medical Research Institute, Tokyo Medical and Dental University
-
- OGURA Atsuo
- RIKEN BioResource Center
Access this Article
Search this Article
Author(s)
-
- OIKAWA Mami OIKAWA Mami
- RIKEN BioResource Center
-
- MATOBA Shogo MATOBA Shogo
- RIKEN BioResource Center
-
- INOUE Kimiko [他] INOUE Kimiko
- RIKEN BioResource Center
-
- KAMIMURA Satoshi
- RIKEN BioResource Center
-
- HIROSE Michiko
- RIKEN BioResource Center
-
- OGONUKI Narumi
- RIKEN BioResource Center
-
- SHIURA Hirosuke
- RIKEN BioResource Center
-
- SUGIMOTO Michihiko
- RIKEN BioResource Center
-
- ABE Kuniya
- RIKEN BioResource Center
-
- ISHINO Fumitoshi
- Department of Epigenetics, Medical Research Institute, Tokyo Medical and Dental University
-
- OGURA Atsuo
- RIKEN BioResource Center
Abstract
In mice, one of the major epigenetic errors associated with somatic cell nuclear transfer (SCNT) is ectopic expression of <i>Xist</i> during the preimplantation period in both sexes. We found that this aberrant <i>Xist</i> expression could be impeded by deletion of <i>Xist</i> from the putative active X chromosome in donor cells. In male clones, it was also found that prior injection of <i>Xist</i>-specific siRNA could significantly improve the postimplantation development of cloned embryos as a result of a significant repression of <i>Xist</i> 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 <i>Xist</i>-specific siRNA at 6–7 h after oocyte activation. RNA FISH analysis revealed that siRNA treatment successfully repressed <i>Xist</i> RNA at the morula stage, as shown by the significant decrease in the number of cloud-type <i>Xist</i> signals in the blastomere nuclei. However, blastomeres with different sizes (from “pinpoint” to “cloud”) and numbers of <i>Xist</i> RNA signals remained within single embryos. After implantation, the dysregulated <i>Xist</i> 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 <i>Xist</i> overexpression in female cloned embryos but failed to rescue them, probably because of an inability to mimic consistent monoallelic <i>Xist</i> expression in these embryos. This could only be achieved in female embryos by applying a gene knockout strategy rather than an siRNA approach.
Journal
-
- Journal of Reproduction and Development
-
Journal of Reproduction and Development 59(3), 231-237, 2013-06-01
THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT
References: 25
-
1
- Phenotypic effects of somatic cell cloning in the mouse
-
OGURA A
Cloning Stem Cells 4, 397-405, 2002
Cited by (1)
-
2
- Mammalian nuclear transfer
-
MEISSNER A
Dev Dyn 235, 2460-2469, 2006
Cited by (1)
-
3
- Significant improvement of mouse cloning technique by treatment with trichostatin A after somatic nuclear transfer
-
KISHIGAMI S
Biochem Biophys Res Commun 340, 183-189, 2006
Cited by (1)
-
4
- Role of histone acetylation in reprogramming of somatic nuclei following nuclear transfer
-
RYBOUCHKIN A
Biol Reprod 74, 1083-1089, 2006
Cited by (1)
-
5
- The histone deacetylase inhibitor scriptaid enhances nascent mRNA production and rescues full-term development in cloned inbred mice
-
VAN THUAN N
Reproduction 138, 309-317, 2009
Cited by (1)
-
6
- Recent advancements in cloning by somatic cell nuclear transfer
-
OGURA A
Philos Trans R Soc Lond B Biol Sci 368, 20110329, 2013
Cited by (1)
-
7
- Impeding Xist expression from the active X chromosome improves mouse somatic cell nuclear transfer
-
INOUE K
Science 330, 496-499, 2010
Cited by (1)
-
8
- Large histone H3 lysine 9 dimethylated chromatin blocks distinguish differentiated from embryonic stem cells
-
WEN B
Nat Genet 41, 246-250, 2009
Cited by (1)
-
9
- RNAi-mediated knockdown of Xist can rescue the impaired postimplantation development of cloned mouse embryos
-
MATOBA S
Proc Natl Acad Sci USA 108, 20621-20626, 2011
Cited by (1)
-
10
- Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei
-
WAKAYAMA T
Nature 394, 369-374, 1998
Cited by (1)
-
11
- Production of male cloned mice from fresh, cultured, and cryopreserved immature Sertoli cells
-
OGURA A
Biol Reprod 62, 1579-1584, 2000
Cited by (1)
-
12
- Culture of preimplantation embryos
-
LAWITTS JA
Methods Enzymol 225, 153-164, 1993
Cited by (1)
-
13
- Latrunculin A can improve the birth rate of cloned mice and simplify the nuclear transfer protocol by gently inhibiting actin polymerization
-
TERASHITA Y
Biol Reprod 86, 180, 2012
Cited by (1)
-
14
- The ground state of embryonic stem cell self-renewal
-
YING QL
Nature 453, 519-523, 2008
Cited by (1)
-
15
- Transcription precedes loss of Xist coating and depletion of H3K27me3 during X-chromosome reprogramming in the mouse inner cell mass
-
WILLIAMS LH
Development 138, 2049-2057, 2011
Cited by (1)
-
16
- Xist-deficient mice are defective in dosage compensation but not spermatogenesis
-
MARAHRENS Y
Genes Dev 11, 156-166, 1997
Cited by (1)
-
17
- Epigenetic dynamics of imprinted X inactivation during early mouse development
-
OKAMOTO I
Science 303, 644-649, 2004
Cited by (1)
-
18
- Choice of random rather than imprinted X inactivation in female embryonic stem cell-derived extra-embryonic cells
-
MURAKAMI K
Development 138, 197-202, 2011
Cited by (1)
-
19
- Failure of extra-embryonic progenitor maintenance in the absence of dosage compensation
-
MUGFORD JW
Development 139, 2130-2138, 2012
Cited by (1)
-
20
- X chromosome inactivation in the cycle of life
-
BARAKAT TS
Development 139, 2085-2089, 2012
Cited by (1)
-
21
- Recent advances in X-chromosome inactivation research
-
POLLEX T
Curr Opin Cell Biol, 2012
Cited by (1)
-
22
- Initiation of epigenetic reprogramming of the X chromosome in somatic nuclei transplanted to a mouse oocyte
-
BAO S
EMBO Rep 6, 748-754, 2005
Cited by (1)
-
23
- Skewed X-inactivation in cloned mice
-
SENDA S
Biochem Biophys Res Commun 321, 38-44, 2004
Cited by (1)
-
24
- X chromosome reactivation and regulation in cloned embryos
-
NOLEN LD
Dev Biol 279, 525-540, 2005
Cited by (1)
-
25
- How to Improve the Success Rate of Mouse Cloning Technology
-
VAN THUAN Nguyen , KISHIGAMI Satoshi , WAKAYAMA Teruhiko
Journal of Reproduction and Development 56(1), 20-30, 2010-02-01
J-STAGE Ichushi Web References (136) Cited by (5)