Successful Mouse Cloning of an Outbred Strain by Trichostatin A Treatment after Somatic Nuclear Transfer

Kishigami, Satoshi; Bui, Hong-Thuy; Wakayama, Sayaka; Tokunaga, Kenzo; Thuan, Nguyen Van; Hikichi, Takafusa; Mizutani, Eiji; Ohta, Hiroshi; Suetsugu, Rinako; Sata, Tetsutaro; Wakayama, Teruhiko

doi:10.1262/jrd.18098

Cited by 140 publications

(93 citation statements)

References 26 publications

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“…Artificially improving the rDNA epigenetic status before or immediately after NT is another option. Recently, several histone deacetylase inhibitors were used to treat the NT embryos to achieve better clone efficiency (52)(53)(54)(55). The histone deacetylase inhibitor can increase the overall histone acetylation level of donor cell chromatins, then make the genes more reachable to maternal factors, therefore, promoting reprogramming.…”

Section: Discussionmentioning

confidence: 99%

rRNA Genes Are Not Fully Activated in Mouse Somatic Cell Nuclear Transfer Embryos

Zheng

Jia

Bou

et al. 2012

Journal of Biological Chemistry

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Background: The nucleolus always shows delayed development and malfunction in somatic cell nuclear transfer (NT) embryos. Results: NT embryos showed low rDNA activity and developmental competence when donor cells with low rDNA activity were used. Conclusion: rDNA reprogramming efficiency in NT embryos was determined by the rDNA activity in donor cells from which they derived. Significance: Developmental potential of NT embryos with rDNA activity in the donor cells was correlated.

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Section: Discussionmentioning

confidence: 99%

rRNA Genes Are Not Fully Activated in Mouse Somatic Cell Nuclear Transfer Embryos

Zheng

Jia

Bou

et al. 2012

Journal of Biological Chemistry

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“…In the mouse, it is well known that the efficiency of SCNT procedures (Wakayama et al 2005, Kishigami et al 2007) and of ESC derivation (Kawase et al 1994, Suzuki et al 1999) is influenced by the genetic background. The observed efficiencies of ESC derivation from our control embryos are similar to those previously reported by Gong et al (2008) in the hybrid B6CBAF1 strain, which was 15.3% for parthenogenetic blastocysts.…”

Section: Discussionmentioning

confidence: 99%

“…These beneficial effects, corroborated in miniature pig SCNT embryos (Miyoshi et al 2010), have recently been questioned by Ono et al (2010), who compared the effects of different HDACis on cloning BD129F1 mice. The discrepancy of results in the mouse species can be probably attributed to strain-specific differences, as not all HDACis are equally effective in all genetic backgrounds (Kishigami et al 2007, Van Thuan et al 2009). With regard to the IE procedure, although Gasparrini et al (2003) previously reported the production of a cloned mouse using cytoplasts prepared by IE, we were unable to obtain live offspring from the IE group.…”

Section: Discussionmentioning

confidence: 99%

Effect of the enucleation procedure on the reprogramming potential and developmental capacity of mouse cloned embryos treated with valproic acid

Costa-Borges

González²,

Santaló³

et al. 2011

Reproduction

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Mouse recipient cytoplasts for somatic cell nuclear transfer (SCNT) are routinely prepared by mechanical enucleation (ME), an invasive procedure that requires expensive equipment and considerable micromanipulation skills. Alternatively, oocytes can be enucleated using chemically assisted (AE) or chemically induced (IE) enucleation methods that are technically simple. In this study, we compared the reprogramming potential and developmental capacity of cloned embryos generated by ME, AE, and IE procedures and treated with the histone deacetylase inhibitor valproic acid. A rapid and almost complete deacetylation of histone H3 lysine 14 in the somatic nucleus followed by an equally rapid and complete re-acetylation after activation was observed after the injection of a cumulus cell nucleus into ME and AE cytoplasts. In contrast, histone deacetylation occurred at a much lower level in IE cytoplasts. Despite these differences, the cloned embryos generated from the three types of cytoplasts developed into blastocysts of equivalent total and inner cell mass mean cell numbers, and the rates of blastocyst formation and embryonic stem cell derivation were similar among the three groups. The cloned embryos produced from ME and AE cytoplasts showed an equivalent rate of full-term development, but no offspring could be obtained from the IE group, suggesting a lower reprogramming capacity of IE cytoplasts. Our results demonstrate the usefulness of AE in mouse SCNT procedures, as an alternative to ME. AE can facilitate oocyte enucleation and avoid the need for expensive microscope optics, or for potentially damaging Hoechst staining and u.v. irradiation, normally required in ME procedures.

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“…The rate of development to morula/blastocyst stage embryos using the nuclei from cells taken from most organs of the frozen mice was very low (Table 1 and Fig. S1c) and not as efficient as when using live cumulus or fibroblast cell nuclei (11)(12)(13). However, cloned embryos derived from brain tissue nuclei developed into morulae/blastocysts (39%) at a rate similar to or even better (Fig.…”

Section: Adaptation Of the Nuclear Transfer Technique For Frozen Deadmentioning

confidence: 95%

Production of healthy cloned mice from bodies frozen at −20°C for 16 years

Wakayama¹,

Ohta²,

Hikichi³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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112

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Cloning animals by nuclear transfer provides an opportunity to preserve endangered mammalian species. However, it has been suggested that the ''resurrection'' of frozen extinct species (such as the woolly mammoth) is impracticable, as no live cells are available, and the genomic material that remains is inevitably degraded. Here we report production of cloned mice from bodies kept frozen at ؊20°C for up to 16 years without any cryoprotection. As all of the cells were ruptured after thawing, we used a modified cloning method and examined nuclei from several organs for use in nuclear transfer attempts. Using brain nuclei as nuclear donors, we established embryonic stem cell lines from the cloned embryos. Healthy cloned mice were then produced from these nuclear transferred embryonic stem cells by serial nuclear transfer. Thus, nuclear transfer techniques could be used to ''resurrect'' animals or maintain valuable genomic stocks from tissues frozen for prolonged periods without any cryopreservation.brain ͉ cloning ͉ cryopreservation ͉ nuclear transfer ͉ reprogramming C urrent animal cloning techniques using nuclear transfer (1) can reconstruct animals from donor cells; clones so produced differ only in their mitochondrial DNA (2). This technology has produced a variety of cloned animals for scientific and commercial purposes. However, it is not known whether this method could be applied to the recovery of animals from frozen extinct species. Although most, if not all, mammalian cells lose their functional integrity if frozen without cryoprotective reagents, it is conceivable that some aspects of genomic information remain intact even in such dead bodies. By contrast, the genomes of spermatozoa are resistant to freezing (3), and even freeze-drying (4) without cryoprotectants. However, it has been difficult to demonstrate this in somatic cells, due to technical difficulties in assessing genomic integrity.One recent paper has reported on the nuclear integrity of dead cells after freeze-thawing by improved techniques (5). This study successfully produced cloned mouse embryos and generated germ line chimerae via nuclear transferred embryonic stem cell (ntES) cells. However, although all donor cells were dead after thawing, the donor cells were frozen in a medium including polyvinylpyrrolidone, which is known to exhibit cryoprotectant effect (6). Freeze-dried cells were also used for these nuclear transfer attempt, and all cells were dead after rehydration (7,8). Although cloned embryos were obtained from those dead cells following nuclear transfer, no cloned offspring were obtained. Moreover, so far all studies of nuclear transfer using ''dead'' material have used isolated single cells frozen in the laboratory using special media. In dead specimens frozen in natural conditions such as permafrost tundra, the cells of tissue will presumably bind strongly to each other and freeze gradually after death due to the large body size. It remains to be shown whether nuclei can be collected from whole bodies frozen without cryopro...

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Successful Mouse Cloning of an Outbred Strain by Trichostatin A Treatment after Somatic Nuclear Transfer

Cited by 140 publications

References 26 publications

rRNA Genes Are Not Fully Activated in Mouse Somatic Cell Nuclear Transfer Embryos

rRNA Genes Are Not Fully Activated in Mouse Somatic Cell Nuclear Transfer Embryos

Effect of the enucleation procedure on the reprogramming potential and developmental capacity of mouse cloned embryos treated with valproic acid

Production of healthy cloned mice from bodies frozen at −20°C for 16 years

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