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CRISPR-Cas9-mediated genome editing in one blastomere of two-cell embryos reveals a novel Tet3 function in regulating neocortical development

Published on Jun 1, 2017in Cell Research17.848
路 DOI :10.1038/cr.2017.58
Lingbo Wang6
Estimated H-index: 6
(CAS: Chinese Academy of Sciences),
Min Yin Li2
Estimated H-index: 2
(CAS: Chinese Academy of Sciences)
+ 19 AuthorsJinsong Li29
Estimated H-index: 29
(CAS: Chinese Academy of Sciences)
Abstract
CRISPR-Cas9-mediated genome editing in one blastomere of two-cell embryos reveals a novel Tet3 function in regulating neocortical development
  • References (42)
  • Citations (9)
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References42
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#1Masakazu Hashimoto (Osaka University)H-Index: 7
#2Yukiko Yamashita (University of Tokushima)H-Index: 2
Last. Tatsuya Takemoto (University of Tokushima)H-Index: 13
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The CRISPR/Cas9 system is a powerful tool for elucidating the roles of genes in a wide variety of organisms including mice. To obtain genetically modified embryos or mice by this method, Cas9 mRNA and sgRNA are usually introduced into zygotes by microinjection or electroporation. However, most mutants generated with this method are genetically mosaic, composed of several types of cells carrying different mutations, which complicates phenotype analysis in founder embryos or mice. To simplify the ...
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#1Takayasu Mikuni (MPG: Max Planck Society)H-Index: 7
#2Jun Nishiyama (MPG: Max Planck Society)H-Index: 9
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A scalable and high-throughput method to identify precise subcellular localization of endogenous proteins is essential for integrative understanding of a cell at the molecular level. Here, we developed a simple and generalizable technique to image endogenous proteins with high specificity, resolution, and contrast in single cells in mammalian brain tissue. The technique, single-cell labeling of endogenous proteins by clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9-mediate...
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Applying CRISPR鈥揅as9 genome editing technologies in safe and reliable ways requires a firm appreciation of the specificity of target-site recognition and cleavage. In this Review the authors discuss various approaches for characterizing off-target effects of CRISPR鈥揅as9 genome editing, how mechanistic knowledge can drive the engineering of more-specific nucleases, and the implications for research and therapeutic applications.
177 CitationsSource
#1Xiaodong Zhu (Rockefeller University)H-Index: 3
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Although mechanisms underlying early steps in cerebellar development are known, evidence is lacking on genetic and epigenetic changes during the establishment of the synaptic circuitry. Using metagene analysis, we report pivotal changes in multiple reactomes of epigenetic pathway genes in cerebellar granule cells (GCs) during circuit formation. During this stage, Tet genes are upregulated and vitamin C activation of Tet enzymes increases the levels of 5-hydroxymethylcytosine (5hmC) at exon start...
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#2Fen Ji (CAS: Chinese Academy of Sciences)H-Index: 6
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Mammalian cortical development is a dynamically and strictly regulated process orchestrated by extracellular signals and intracellular mechanisms. Recent studies show that epigenetic regulation serves as, at least in part, interfaces between genes and the environment, and also provides insight into the molecular and cellular bases of early embryonic cortical development. It is becoming increasingly clear that epigenetic regulation of cortical development occurs at multiple levels and that compre...
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#1Huimei Yu (Johns Hopkins University)H-Index: 4
#2Yijing Su (Johns Hopkins University)H-Index: 18
Last. Hongjun Song (Johns Hopkins University)H-Index: 82
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The cellular function of active DNA demethylation in neurons is not well understood. Here, Song and colleagues show that synaptic activity modulates Tet3 signaling, which in turn regulates glutamatergic synaptic transmission and synaptic scaling. Their work identifies Tet3 as a synaptic activity sensor to epigenetically regulate fundamental properties and meta-plasticity of neurons via active DNA demethylation.
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#1Ting Li (CAS: Chinese Academy of Sciences)H-Index: 8
#2Dehua Yang (CAS: Chinese Academy of Sciences)H-Index: 21
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5-Hydroxymethylcytosine (5hmC), converted from 5-methylcytocine (5mC) by Tet family of dioxygenases (Tet1, Tet2, and Tet3), is enriched in the embryonic stem cells (ESCs) and in the brain. However, the role of 5hmC and Tet family in the process of ESC differentiation especially neuronal differentiation remains elusive. Here, we showed the evidence that Tet3 is critical in neural progenitor cell (NPC) maintenance and terminal differentiation of neurons. We found that Tet3 expression is basically ...
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#1Lukasz SwiechH-Index: 4
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Probing gene function in the mammalian brain can be greatly assisted with methods to manipulate the genome of neurons in vivo. The clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated endonuclease (Cas)9 from Streptococcus pyogenes (SpCas9)1 can be used to edit single or multiple genes in replicating eukaryotic cells, resulting in frame-shifting insertion/deletion (indel) mutations and subsequent protein depletion. Here, we delivered SpCas9 and guide RNAs using adeno-a...
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#1Jennifer A. Doudna (University of California, Berkeley)H-Index: 98
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The advent of facile genome engineering using the bacterial RNA-guided CRISPR-Cas9 system in animals and plants is transforming biology. We review the history of CRISPR (clustered regularly interspaced palindromic repeat) biology from its initial discovery through the elucidation of the CRISPR-Cas9 enzyme mechanism, which has set the stage for remarkable developments using this technology to modify, regulate, or mark genomic loci in a wide variety of cells and organisms from all three domains of...
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Abstract Epigenetic modifications of the genome play important roles in controlling gene transcription thus regulating several molecular and cellular processes. A novel epigenetic modification 鈥 5-hydroxymethylcytosine (5hmC) 鈥 has been recently described and attracted a lot of attention due to its possible involvement in the active DNA demethylation mechanism. TET enzymes are dioxygenases capable of oxidizing the methyl group of 5-methylcytosines (5mC) and thus converting 5mC into 5hmC. Althoug...
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TET3 is a member of the ten-eleven translocation (TET) family of enzymes which oxidize 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC). Tet3 is highly expressed in the brain, where 5hmC levels are most abundant. In adult mice, we observed that TET3 is present in mature neurons and oligodendrocytes but is absent in astrocytes. To investigate the function of TET3 in adult postmitotic neurons, we crossed Tet3 floxed mice with a neuronal Cre-expressing mouse line, Camk2a-CreERT2, obtainin...
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