globalchange  > 气候变化与战略
DOI: 10.1073/pnas.1817703116
论文题名:
DNA methylation analysis and editing in single mammalian oocytes
作者: Wei Y.; Lang J.; Zhang Q.; Yang C.-R.; Zhao Z.-A.; Zhang Y.; Du Y.; Sun Y.
刊名: Proceedings of the National Academy of Sciences of the United States of America
ISSN: 0027-8424
出版年: 2019
卷: 116, 期:20
起始页码: 9883
结束页码: 9892
语种: 英语
英文关键词: DNA methylation ; Epigenetic inheritance ; Oocyte
Scopus关键词: bisulfite ; DNA methyltransferase 3A ; adult ; animal cell ; animal experiment ; animal model ; animal tissue ; Article ; controlled study ; CRISPR-CAS9 system ; DNA determination ; DNA flanking region ; DNA methylation ; DNA sequence ; embryo ; female ; gene editing ; gene targeting ; genetic linkage ; genetic manipulation ; genome imprinting ; happy puppet syndrome ; male ; mammal cell ; microinjection ; mouse ; newborn ; nonhuman ; oocyte ; parthenogenesis ; phenotype ; polar body ; priority journal ; process optimization ; single cell analysis ; animal ; C57BL mouse ; genetic engineering ; metabolism ; procedures ; single cell analysis ; Animals ; DNA Methylation ; Female ; Genetic Engineering ; Mice, Inbred C57BL ; Polar Bodies ; Single-Cell Analysis
英文摘要: Mammalian oocytes carry specific nongenetic information, including DNA methylation to the next generation, which is important for development and disease. However, evaluation and manipulation of specific methylation for both functional analysis and therapeutic purposes remains challenging. Here, we demonstrate evaluation of specific methylation in single oocytes from its sibling first polar body (PB1) and manipulation of specific methylation in single oocytes by microinjection-mediated dCas9-based targeted methylation editing. We optimized a single-cell bisulfite sequencing approach with high efficiency and demonstrate that the PB1 carries similar methylation profiles at specific regions to its sibling oocyte. By bisulfite sequencing of a single PB1, the methylation information regarding agouti viable yellow (Avy)-related coat color, as well as imprinting linked parthenogenetic development competency, in a single oocyte can be efficiently evaluated. Microinjection-based dCas9-Tet/Dnmt–mediated methylation editing allows targeted manipulation of specific methylation in single oocytes. By targeted methylation editing, we were able to reverse Avy-related coat color, generate full-term development of bimaternal mice, and correct familial Angelman syndrome in a mouse model. Our work will facilitate the investigation of specific methylation events in oocytes and provides a strategy for prevention and correction of maternally transmitted nongenetic disease or disorders. © 2019 National Academy of Sciences. All rights reserved.
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资源类型: 期刊论文
标识符: http://119.78.100.158/handle/2HF3EXSE/163556
Appears in Collections:气候变化与战略

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作者单位: Wei, Y., Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200135, China, Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai, 200135, China; Lang, J., Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200135, China, Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai, 200135, China, Center for Reproductive Medicine, Shandong University, Jinan, 250021, China, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Jinan, 250021, China; Zhang, Q., Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200135, China, Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai, 200135, China, Center for Reproductive Medicine, Shandong University, Jinan, 250021, China, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Jinan, 250021, China; Yang, C.-R., Center for Reproductive Sciences, University of California, San Francisco, CA 94143, United States; Zhao, Z.-A., Institute for Cardiovascular Science, Soochow University, Suzhou, 215000, China; Zhang, Y., Broad Institute of MIT and Harvard, Cambridge, 02142, United States; Du, Y., Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200135, China, Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai, 200135, China; Sun, Y., Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200135, China, Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai, 200135, China

Recommended Citation:
Wei Y.,Lang J.,Zhang Q.,et al. DNA methylation analysis and editing in single mammalian oocytes[J]. Proceedings of the National Academy of Sciences of the United States of America,2019-01-01,116(20)
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