Inter-homologue repair in fertilized human eggs?

Published on Aug 1, 2018in Nature 41.58
· DOI :10.1038/s41586-018-0379-5
Dieter Egli34
Estimated H-index: 34
(Columbia University),
Michael V. Zuccaro1
Estimated H-index: 1
(Columbia University)
+ 3 AuthorsMaria Jasin77
Estimated H-index: 77
(Memorial Sloan Kettering Cancer Center)
Abstract
Many human diseases have an underlying genetic component. The development and application of methods to prevent the inheritance of damaging mutations through the human germline could have significant health benefits, and currently include preimplantation genetic diagnosis and carrier screening. Ma et al. take this a step further by attempting to remove a disease mutation from the human germline through gene editing (1). They assert the following advances: (i) the correction of a pathogenic gene mutation responsible for hypertrophic cardiomyopathy in human embryos using CRISPR-Cas9 and (ii) the avoidance of mosaicism in edited embryos. In the case of correction, the authors conclude that repair using the homologous chromosome was as or more frequent than mutagenic nonhomologous end-joining (NHEJ). Their conclusion is significant, if validated, because such a self-repair mechanism would allow gene correction without the introduction of a repair template. While the authors analyses relied on the failure to detect mutant alleles, here we suggest approaches to provide direct evidence for inter-homologue recombination and discuss other events consistent with the data. We also review the biological constraints on inter-homologue recombination in the early embryo. (1) Ma, H. et al. Correction of a pathogenic gene mutation in human embryos. Nature, doi:10.1038/nature23305 (2017).
  • References (16)
  • Citations (18)
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References16
Published on Jun 7, 2007in Nature 41.58
Dieter Egli34
Estimated H-index: 34
(Harvard University),
Jacqueline Rosains3
Estimated H-index: 3
(Harvard University)
+ 1 AuthorsKevin Eggan52
Estimated H-index: 52
(Harvard University)
It is generally believed that unfertilized oocytes are required for successful somatic cell nuclear transfer, but this paper demonstrates that reprogramming activities persist after fertilization. These findings have important implications for understanding the nature of the nuclear reprogramming activities, the biology of cloned animals and the ongoing efforts to derive patient specific human embryonic stem cell lines.
261 Citations Source Cite
Published on Apr 1, 2004in Nature Genetics 27.13
Alec J. Jeffreys67
Estimated H-index: 67
(University of Leicester),
Celia A. May17
Estimated H-index: 17
(University of Leicester)
Erratum: Intense and highly localized gene conversion activity in human meiotic crossover hot spots
228 Citations Source Cite
Published on Jan 1, 1995in Human Reproduction 4.99
K.M. Sultan1
Estimated H-index: 1
(Cornell University),
Santiago Munné73
Estimated H-index: 73
(Cornell University)
+ 2 AuthorsJacques Cohen75
Estimated H-index: 75
(Cornell University)
Uni-pronuclear embryos (n=42) were analysed by fluorescence in-situ hybridization (FISH) with two to four chromosome pair-specific probes. Half of these embryos resulted from conventional insemination and half from intracytoplasmic sperm injection (ICSI). The majority of unipronuclear embryos from conventional insemination were normally diploid (61.9%) whereas only 9.5% of unipronuclear ICSI embryos (P<0.001) were diploid. In addition, a significantly higher number of uni-pronuclear embryos from...
58 Citations Source Cite
Published on Oct 1, 2014in Nature Genetics 27.13
Francesca Cole17
Estimated H-index: 17
,
Frédéric Baudat22
Estimated H-index: 22
+ 3 AuthorsMaria Jasin10
Estimated H-index: 10
(Memorial Sloan Kettering Cancer Center)
Scott Keeney, Bernard de Massy, Maria Jasin and colleagues report a method to perform tetrad analysis (analysis of all four chromatids from a single meiosis) in mouse and analyzed two recombination hotspots in mouse oocytes and spermatocytes. They show that gene conversion frequently spares the binding site of the hotspot-specifying protein PRDM9.
46 Citations Source Cite
Published on Sep 1, 2011in Nature Communications 12.35
Dieter Egli34
Estimated H-index: 34
(New York Stem Cell Foundation),
Alice E. Chen11
Estimated H-index: 11
(Harvard University)
+ 11 AuthorsWilliam G. Kearns1
Estimated H-index: 1
The generation of human cell lines using somatic cell nuclear transfer has been difficult to achieve. In this study, Egli et al. show that while mouse eggs reprogram somatic cells within hours, human eggs arrest after nuclear transfer which may be due to a lack of genome transcription.
54 Citations Source Cite
Published on May 1, 2015in Scientific Reports 4.12
Toru Suzuki5
Estimated H-index: 5
,
Maki Asami3
Estimated H-index: 3
,
Anthony C.F. Perry32
Estimated H-index: 32
Mammalian genomes can be edited by injecting pronuclear embryos with Cas9 cRNA and guide RNA (gRNA) but it is unknown whether editing can also occur during the onset of embryonic development, prior to pronuclear embryogenesis. We here report Cas9-mediated editing during sperm-induced meiotic exit and the initiation of development. Injection of unfertilized, mouse metaphase II (mII) oocytes with Cas9 cRNA, gRNA and sperm enabled efficient editing of transgenic and native alleles. Pre-loading oocy...
22 Citations Source Cite
Published on Jan 14, 2015in PLOS ONE 2.77
Bijal A. Parikh4
Estimated H-index: 4
(Washington University in St. Louis),
Diana L. Beckman8
Estimated H-index: 8
(Washington University in St. Louis)
+ 2 AuthorsWayne M. Yokoyama72
Estimated H-index: 72
(Washington University in St. Louis)
The bacterial CRISPR-Cas9 system has been adapted for use as a genome editing tool. While several recent reports have indicated that successful genome editing of mice can be achieved, detailed phenotypic and molecular analyses of the mutant animals are limited. Following pronuclear micro-injection of fertilized eggs with either wild-type Cas9 or the nickase mutant (D10A) and single or paired guide RNA (sgRNA) for targeting of the tyrosinase (Tyr) gene, we assessed genome editing in mice using ra...
29 Citations Source Cite
Published on Jun 1, 2003in Human Reproduction 4.99
Suresh Kattera6
Estimated H-index: 6
,
Christopher Chen6
Estimated H-index: 6
Microsurgical enucleation of a single pronucleus from each of three tripronuclear zygotes was performed and the embryos were transferred to a 38-year-old woman on day 3 after fertilization. A normal healthy baby boy was born at 38 weeks and 4 days gestation, demonstrating that with polyspermic fertilization, removal of the extra male pronucleus allows the zygote to develop normally.
29 Citations Source Cite
Published on Jan 15, 2003in Molecular and Cellular Biology 3.81
Jeremy M. Stark25
Estimated H-index: 25
,
Maria Jasin8
Estimated H-index: 8
Loss of heterozygosity (LOH) is a common genetic alteration in tumors and often extends several megabases to encompass multiple genetic loci or even whole chromosome arms. Based on marker and karyotype analysis of tumor samples, a significant fraction of LOH events appears to arise from mitotic recombination between homologous chromosomes, reminiscent of recombination during meiosis. As DNA double-strand breaks (DSBs) initiate meiotic recombination, a potential mechanism leading to LOH in mitoti...
88 Citations Source Cite
Published on Apr 15, 2013in Genes & Development 9.46
Liisa Kauppi12
Estimated H-index: 12
(University of Helsinki),
Marco Barchi15
Estimated H-index: 15
(University of Rome Tor Vergata)
+ 3 AuthorsScott Keeney46
Estimated H-index: 46
(Memorial Sloan Kettering Cancer Center)
Different organisms display widely different numbers of the programmed double-strand breaks (DSBs) that initiate meiotic recombination (e.g., hundreds per meiocyte in mice and humans vs. dozens in nematodes), but little is known about what drives these species-specific DSB set points or the regulatory pathways that control them. Here we examine male mice with a lowered dosage of SPO11, the meiotic DSB catalyst, to gain insight into the effect of reduced DSB numbers on mammalian chromosome dynami...
86 Citations Source Cite
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Cited By18
Published on Oct 10, 2017in PLOS Pathogens 6.16
Spencer S. Gang4
Estimated H-index: 4
(University of California, Los Angeles),
Michelle L. Castelletto6
Estimated H-index: 6
(University of California, Los Angeles)
+ 5 AuthorsElissa A. Hallem16
Estimated H-index: 16
(University of California, Los Angeles)
Parasitic nematodes infect over 1 billion people worldwide and cause some of the most common neglected tropical diseases. Despite their prevalence, our understanding of the biology of parasitic nematodes has been limited by the lack of tools for genetic intervention. In particular, it has not yet been possible to generate targeted gene disruptions and mutant phenotypes in any parasitic nematode. Here, we report the development of a method for introducing CRISPR-Cas9-mediated gene disruptions in ...
15 Citations Source Cite
Published on Mar 1, 2018in Clinical Chemistry 8.64
Françoise Baylis23
Estimated H-index: 23
(Dalhousie University)
The anticipated benefits of human gene editing with the use of CRISPR (clustered interspaced short palindromic repeats)-Cas9 are both familiar and contested. First and foremost is the expectation of cures for blood disorders, lung diseases, cancers, and other maladies as clinician-scientists master various insertion, disruption, and deletion techniques. In addition to these potential therapeutic benefits, for some, there are the potential benefits of human enhancement as investigators learn to m...
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Published on May 7, 2018in Molecular Reproduction and Development 3.11
Lichun Tang1
Estimated H-index: 1
(Protein Sciences),
Yanting Zeng1
Estimated H-index: 1
+ 4 AuthorsPumin Zhang35
Estimated H-index: 35
(Protein Sciences)
Source Cite
Published on Apr 1, 2018in JACC: Basic to Translational Science
Alexandra Dainis1
Estimated H-index: 1
(Stanford University),
Euan A. Ashley54
Estimated H-index: 54
Summary Precision medicine strives to delineate disease using multiple data sources—from genomics to digital health metrics—in order to be more precise and accurate in our diagnoses, definitions, and treatments of disease subtypes. By defining disease at a deeper level, we can treat patients based on an understanding of the molecular underpinnings of their presentations, rather than grouping patients into broad categories with one-size-fits-all treatments. In this review, the authors examine how...
1 Citations Source Cite
Published on Aug 1, 2018in Journal of Assisted Reproduction and Genetics 2.79
A. Hershlag2
Estimated H-index: 2
(Hofstra University),
Sara L. Bristow1
Estimated H-index: 1
(Hofstra University)
The rapid development of gene-editing technologies has led to an exponential rise in both basic and translational research initiatives studying molecular processes and investigating possible clinical applications. Early experiments using genome editing to study human embryo development have contradicted findings in studies on model organisms. Additionally, a series of four experiments over the past 2 years set out to investigate the possibilities of introducing genetic modifications to human emb...
1 Citations Source Cite
Published on Sep 1, 2018in British Medical Bulletin 3.36
Andy Greenfield27
Estimated H-index: 27
(Medical Research Council)
2 Citations Source Cite
Published on Aug 8, 2018in Nature 41.58
Ewen Callaway Ec17
Estimated H-index: 17
Researchers provide more evidence for their landmark claim that gene editing rid embryos of a disease mutation — but scientists are still arguing over the results. Researchers provide more evidence for their landmark claim that gene editing rid embryos of a disease mutation — but scientists are still arguing over the results.
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Published on Dec 1, 2018in Seminars in Perinatology 2.88
Natalie Kofler2
Estimated H-index: 2
(Yale University),
Katherine L. Kraschel (Yale University)
Abstract CRISPR gene editing is poised to transform the therapeutic landscape for diseases of genetic origin. The ease and agility by which CRISPR can make specific changes to DNA holds great promise not only for the treatment of heritable diseases, but also their prevention through germline editing. CRISPR-based therapeutic strategies are currently under development for numerous monogenic diseases. These strategies range from proof of concept studies demonstrating pre-fertilization gamete editi...
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Published on Dec 1, 2018in Seminars in Perinatology 2.88
Fani Memi4
Estimated H-index: 4
(University College London),
Aglaia Ntokou1
Estimated H-index: 1
(Yale University),
Irinna Papangeli7
Estimated H-index: 7
(Yale University)
Abstract Gene therapy carries the potential to treat more than 10,000 human monogenic diseases and benefit an even greater number of complex polygenic conditions. The repurposing of CRISPR/Cas9, an ancient bacterial immune defense system, into a gene-editing technology has armed researchers with a revolutionary tool for gene therapy. However, as the breadth of research and clinical applications of this technology continues to expand, outstanding technical challenges and ethical considerations wi...
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Published on Dec 1, 2018in Synthetic and Systems Biotechnology
Huayi Liu1
Estimated H-index: 1
(Sichuan University),
Lian Wang1
Estimated H-index: 1
(Sichuan University),
Yunzi Luo12
Estimated H-index: 12
(Sichuan University)
Abstract Since 2013, the CRISPR-based bacterial antiviral defense systems have revolutionized the genome editing field. In addition to genome editing, CRISPR has been developed as a variety of tools for gene expression regulations, live cell chromatin imaging, base editing, epigenome editing, and nucleic acid detection. Moreover, in the context of further boosting the usability and feasibility of CRISPR systems, novel CRISPR systems and engineered CRISPR protein mutants have been explored and st...
1 Citations Source Cite