Nuclear genome transfer in human oocytes eliminates mitochondrial DNA variants

Paull, Daniel; Emmanuele, Valentina; Weiss, Keren A.; Treff, Nathan R.; Stewart, Latoya A.; Hua, Haiqing; Zimmer, Matthew; Kahler, David J.; Goland, Robin; Noggle, Scott; Prosser, Robert; Hirano, Michio; Sauer, Mark V.; Egli, Dieter

doi:10.1038/nature11800

Cited by 232 publications

(228 citation statements)

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“…Obtained (pro)nuclear intensities were used to calculate relative intensities (paternal PN/maternal PN for zygote, and nucleus/cytosol for 2-cell embryo [42]). Statistical analysis for embryonic survival rates was performed using the chi-square test [43,44]. For other experiments, detailed information about quantification and statistical analyses is described in the Supplementary Materials and Methods.…”

Section: Data Quantification and Statistical Analysismentioning

confidence: 99%

Paternal H3K4 methylation is required for minor zygotic gene activation and early mouse embryonic development

et al. 2015

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Epigenetic modifications, such as DNA methylation and histone modifications, are dynamically altered predominantly in paternal pronuclei soon after fertilization. To identify which histone modifications are required for early embryonic development, we utilized histone K-M mutants, which prevent endogenous histone methylation at the mutated site. We prepared four single K-M mutants for histone H3.3, K4M, K9M, K27M, and K36M, and demonstrate that overexpression of H3.3 K4M in embryos before fertilization results in developmental arrest, whereas overexpression after fertilization does not affect the development. Furthermore, loss of H3K4 methylation decreases the level of minor zygotic gene activation (ZGA) predominantly in the paternal pronucleus, and we obtained similar results from knockdown of the H3K4 methyltransferase Mll3/4. We therefore conclude that H3K4 methylation, likely established by Mll3/4 at the early pronuclear stage, is essential for the onset of minor ZGA in the paternal pronucleus, which is necessary for subsequent preimplantation development in mice.

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Section: Data Quantification and Statistical Analysismentioning

confidence: 99%

Paternal H3K4 methylation is required for minor zygotic gene activation and early mouse embryonic development

et al. 2015

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“…These approaches are now almost exclusively oriented on the elimination of mutated mitochondrial DNA (mtDNA), that can cause some devastating diseases [29]. It has been demonstrated quite recently in humans that the transfer of pronuclei or spindles, isolated from the cytoplasm containing mutated mtDNA, into healthy cytoplasts seems to be very promising and healthy looking blastocysts were obtained [30][31][32][33].…”

Section: Discussionmentioning

confidence: 99%

The ups and downs of somatic cell nucleus transfer (SCNT) in humans

Fulka

Langerova

Loi

et al. 2013

J Assist Reprod Genet

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Achieving successful somatic cell nuclear transfer (SCNT) in the human and subhuman primate relative to other mammals has been questioned for a variety of technical and logistical issues. Here we summarize the gradual evolution of SCNT technology from the perspective of oocyte quality and cell cycle status that has recently led to the demonstration of feasibility in the human for deriving chromosomally normal stem cells lines. With these advances in hand, prospects for therapeutic cloning must be entertained in a conscientious, rigorous, and timely fashion before broad spectrum clinical applications are undertaken.Keywords Nucleus . Oocyte . Nucleus transfer Simply stated, SCNT in mammals should in practice be a very straightforward technique. First, cytoplasts are prepared by enucleating the metaphase II stage oocytes, i.e. metaphase II chromosomes are removed from the oocyte. Second, the selected nucleus is introduced into the cytoplast either by direct injection or by induced cell fusion. The reconstructed SCNT products are then activated in ways mimicking fertilization and if everything goes well cloned embryos develop [6].Logically, since following the birth of Dolly [7], and the production of additional clones in other mammalian species, there is ample confirmation that Dolly was not an exceptional case, prompting widespread discussion regarding the pros and cons of human SCNT. Given this prospect, it has been commonly accepted and broadly emphasized that the production of cloned human individuals must be banned (reproductive cloning). On the other hand, the production of embryos from which patient compatible embryonic stem cells could be obtained seemed to be acceptable, at least in some countries (therapeutic cloning). However, once the discovery of induced pluripotent stem cell production (iPSC) was realized by the work of Takahashi and Yamanaka [see 8 for review], illustrating that differentiated or even terminally differentiated cells can be converted (dedifferentiated) into a pluripotent state by induced overexpression of four factors (Oct 4, Sox 2, Klf 4, c-Myc; OSKM), greater resistance against the use of SCNT in humans has emerged, including the prospect of therapeutic cloning.Although many papers on modifications of the original iPS technology seeking improved efficiency and safety have been published, it should be emphasized that our understanding of the mechanisms by which iPSC elicits dedifferentiation remain woefully incomplete. Instead, it is commonly accepted that the oocyte cytoplasmic milieu is the most direct means to obtaining reprogramming of a differentiated somatic cell nucleus [9,10]. This fact-the quality of the oocyte cytoplasmic milieu-may well underscore the reason that while research on human SCNT continues, reports are rare in this species relative to the many papers published in other mammals. Birth of DollyThe work that led to the birth of Dolly was antedated by many years of experiments using laboratory and domestic animals to test many methodological combinations ...

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“…Unlike nuclear disorders, germline gene therapy is being considered for mitochondrial disorders and, indeed, significant strides have been made recently, with reports indicating that successful transfer of the nuclear genetic material from human oocytes carrying mutant mtDNA to enucleated recipient oocytes containing wild type mitochondrial genomes has been achieved [83,84] (Figure 2). Of note, in previous studies, genome exchange between oocytes of nonhuman primates resulted in production of healthy offspring [85] and pronuclear transfer between fertilised human zygotes had been achieved [86].…”

Section: Trends In Geneticsmentioning

confidence: 99%

“…In essence, the approach involves using an enucleated egg from a donor female into which the nucleus from the mother's egg is inserted, followed by fertilisation by the father's sperm of the 'hybrid' egg, whose mtDNA material originates from the donor female and nuclear material from the mother. Significant strides have been made recently towards this objective [83][84][85][86]. Reproduced with kind permission from Shoukhrat Mitalipov, Oregon Health & Science University.…”

Section: The Mitochondriamentioning

confidence: 99%