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One-step generation of complete gene knockout mice and monkeys by CRISPR/Cas9-mediated gene editing with multiple sgRNAs

Published on Jul 1, 2017in Cell Research17.848
路 DOI :10.1038/cr.2017.81
Erwei Zuo6
Estimated H-index: 6
(CAS: Chinese Academy of Sciences),
Yijun Cai8
Estimated H-index: 8
(CAS: Chinese Academy of Sciences)
+ 26 AuthorsHui Yang19
Estimated H-index: 19
(CAS: Chinese Academy of Sciences)
Sources
Abstract
One-step generation of complete gene knockout mice and monkeys by CRISPR/Cas9-mediated gene editing with multiple sgRNAs
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  • References (37)
  • Citations (47)
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References37
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#2Aida Platero-Luengo (Salk Institute for Biological Studies)H-Index: 3
Last. Juan Carlos Izpisua Belmonte (Salk Institute for Biological Studies)H-Index: 84
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Interspecies blastocyst complementation enables organ-specific enrichment of xenogenic pluripotent stem cell (PSC) derivatives. Here, we establish a versatile blastocyst complementation platform based on CRISPR-Cas9-mediated zygote genome editing and show enrichment of rat PSC-derivatives in several tissues of gene-edited organogenesis-disabled mice. Besides gaining insights into species evolution, embryogenesis, and human disease, interspecies blastocyst complementation might allow human organ ...
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#1Hai-Qiang DaiH-Index: 2
#2Bang-An WangH-Index: 3
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Inactivation of three Tet genes in mice leads to gastrulation phenotypes similar to those in embryos with increased Nodal signalling, revealing a functional redundancy of Tet genes and showing balanced and dynamic DNA methylation and demethylation is crucial to regulate key signalling pathways in early body plan formation.
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#1Masakazu Hashimoto (Osaka University)H-Index: 7
#2Yukiko Yamashita (University of Tokushima)H-Index: 2
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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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#1Charles Jennings C (MIT: Massachusetts Institute of Technology)H-Index: 7
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Given recent advances in genome engineering technology like CRISPR and the difficulty of modeling human diseases in rodents, transgenic nonhuman primates may be used to develop etiologically relevant models of disease. This perspective by Guoping Feng et al. highlights the technological advances, potential challenges and opportunities these models present to furthering our understanding of disease.
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A CRISPR/Cas9 genome editing framework has been developed that allows controlled introduction of mono- and bi-allelic sequence changes, and is used to generate induced human pluripotent stem cells with heterozygous and homozygous dominant mutations in amyloid precursor protein and presenilin 1 that have been associated with early onset Alzheimer鈥檚 disease.
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Dioxygenases of the TET (Ten-Eleven Translocation) family produce oxidized methylcytosines, intermediates in DNA demethylation, as well as new epigenetic marks. Here we show data suggesting that TET proteins maintain the consistency of gene transcription. Embryos lacking Tet1 and Tet3 (Tet1/3 DKO) displayed a strong loss of 5-hydroxymethylcytosine (5hmC) and a concurrent increase in 5-methylcytosine (5mC) at the eight-cell stage. Single cells from eight-cell embryos and individual embryonic day ...
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Some reports demonstrated successful genome editing in pigs by one-step zygote microinjection of mRNA of CRISPR/Cas9-related components. Given the relatively long gestation periods and the high cost of housing, the establishment of a single blastocyst-based assay for rapid optimization of the above system is required. As a proof-of-concept, we attempted to disrupt a gene (GGTA1) encoding the 伪-1,3-galactosyltransferase that synthesizes the 伪-Gal epitope using parthenogenetically activated porcin...
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CRISPR/Cas9 has been used to genetically modify genomes in a variety of species, including non-human primates. Unfortunately, this new technology does cause mosaic mutations, and we do not yet know whether such mutations can functionally disrupt the targeted gene or cause the pathology seen in human disease. Addressing these issues is necessary if we are to generate large animal models of human diseases using CRISPR/Cas9. Here we used CRISPR/Cas9 to target the monkey dystrophin gene to create mu...
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A genome editing technique based on the clustered regularly interspaced short palindromic repeats (CRISPR)鈥揳ssociated endonuclease Cas9 enables efficient modification of genes in various cell types, including neurons. However, neuronal ensembles even in the same brain region are not anatomically or functionally uniform but divide into distinct subpopulations. Such heterogeneity requires gene editing in specific neuronal populations. We developed a CRISPR-SaCas9 system鈥揵ased technique, and its co...
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ABSTRACT Owing to their high similarity to humans, non-human primates (NHPs) provide an exceedingly suitable model for the study of human disease. In this Review, we summarize the history of transgenic NHP models and the progress of CRISPR/Cas9-mediated genome editing in NHPs, from the first proof-of-principle green fluorescent protein-expressing monkeys to sophisticated NHP models of human neurodegenerative disease that accurately phenocopy several complex disease features. We discuss not only ...
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