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Generation of gene-modified cynomolgus monkey via Cas9/RNA-mediated gene targeting in one-cell embryos.

Published on Feb 1, 2014in Cell 31.40
· DOI :10.1016/j.cell.2014.01.027
Yuyu Niu15
Estimated H-index: 15
,
Bin Shen14
Estimated H-index: 14
(National Resource Center)
+ 25 AuthorsJiahao Sha33
Estimated H-index: 33
(Nanjing Medical University)
Abstract
Summary Monkeys serve as important model species for studying human diseases and developing therapeutic strategies, yet the application of monkeys in biomedical researches has been significantly hindered by the difficulties in producing animals genetically modified at the desired target sites. Here, we first applied the CRISPR/Cas9 system, a versatile tool for editing the genes of different organisms, to target monkey genomes. By coinjection of Cas9 mRNA and sgRNAs into one-cell-stage embryos, we successfully achieve precise gene targeting in cynomolgus monkeys. We also show that this system enables simultaneous disruption of two target genes ( Ppar-γ and Rag1 ) in one step, and no off-target mutagenesis was detected by comprehensive analysis. Thus, coinjection of one-cell-stage embryos with Cas9 mRNA and sgRNAs is an efficient and reliable approach for gene-modified cynomolgus monkey generation.
  • References (25)
  • Citations (584)
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References25
Newest
Published on Jan 1, 2014in Cell Research 15.39
Yuanwu Ma6
Estimated H-index: 6
,
Xu Zhang7
Estimated H-index: 7
+ 6 AuthorsLianfeng Zhang6
Estimated H-index: 6
114 Citations Source Cite
Jiankui Zhou5
Estimated H-index: 5
(National Resource Center),
Bin Shen14
Estimated H-index: 14
(National Resource Center)
+ 9 AuthorsBian Hu7
Estimated H-index: 7
(National Resource Center)
Taking advantage of the multiplexable genome engineering feature of the CRISPR/Cas9 system, we sought to generate different kinds of immunodeficient mouse strains by embryo co-microinjection of Cas9 mRNA and multiple sgRNAs targeting mouse B2m, Il2rg, Prf1, Prkdc, and Rag1. We successfully achieved multiple gene modifications, fragment deletion, double knockout of genes localizing on the same chromosome, and got different kinds of immunodeficient mouse models with different heritable genetic mod...
75 Citations Source Cite
Published on Nov 6, 2013in Nature 41.58
Helen Shen7
Estimated H-index: 7
16 Citations Source Cite
Published on Oct 1, 2013in Nature Methods 26.92
Prashant Mali29
Estimated H-index: 29
,
Kevin M. Esvelt16
Estimated H-index: 16
,
George M Church G M135
Estimated H-index: 135
This Perspective describes current and prospective advances in genome engineering made possible with the CRISPR-Cas9 system.
727 Citations Source Cite
Published on Sep 1, 2013in Cell 31.40
F. Ann Ran9
Estimated H-index: 9
,
Patrick Hsu15
Estimated H-index: 15
+ 8 AuthorsYiyun Zhang94
Estimated H-index: 94
Summary Targeted genome editing technologies have enabled a broad range of research and medical applications. The Cas9 nuclease from the microbial CRISPR-Cas system is targeted to specific genomic loci by a 20 nt guide sequence, which can tolerate certain mismatches to the DNA target and thereby promote undesired off-target mutagenesis. Here, we describe an approach that combines a Cas9 nickase mutant with paired guide RNAs to introduce targeted double-strand breaks. Because individual nicks in ...
1,705 Citations Source Cite
Published on Sep 1, 2013in Nature Biotechnology 35.72
Vikram Pattanayak11
Estimated H-index: 11
,
Steven Lin24
Estimated H-index: 24
+ 3 AuthorsDavid R. Liu69
Estimated H-index: 69
In vitro selection and high-throughput sequencing measure the sequence specificity of cleavage by gRNA-Cas9 complexes.
756 Citations Source Cite
Published on Sep 1, 2013in Nature Biotechnology 35.72
Yanfang Fu14
Estimated H-index: 14
(Chinese Academy of Sciences),
Jennifer A. Foden4
Estimated H-index: 4
(Chinese Academy of Sciences)
+ 4 AuthorsJeffry D. Sander4
Estimated H-index: 4
(Chinese Academy of Sciences)
CRISPR RNA-guided endonucleases (RGENs) have rapidly emerged as a facile and efficient platform for genome editing. Here, we use a human cell-based reporter assay to characterize off-target cleavage of Cas9-based RGENs. We find that single and double mismatches are tolerated to varying degrees depending on their position along the guide RNA (gRNA)-DNA interface. We readily detected off-target alterations induced by four out of six RGENs targeted to endogenous loci in human cells by examination o...
1,599 Citations Source Cite
Published on Sep 1, 2013in Nature Biotechnology 35.72
Patrick Hsu15
Estimated H-index: 15
,
Douglas Scott96
Estimated H-index: 96
+ 11 AuthorsOphir Shalem18
Estimated H-index: 18
Analyses of the determinants of the specificity of Cas9 nuclease provide rules for selecting optimal target sites.
2,006 Citations Source Cite
Published on Aug 1, 2013in Nature Biotechnology 35.72
Dali Li29
Estimated H-index: 29
,
Zhongwei Qiu5
Estimated H-index: 5
+ 9 AuthorsXiaoling Lu12
Estimated H-index: 12
395 Citations Source Cite
Published on Aug 1, 2013in Nature Biotechnology 35.72
Wei Li41
Estimated H-index: 41
,
Fei Teng8
Estimated H-index: 8
+ 1 AuthorsQi Zhou51
Estimated H-index: 51
Simultaneous generation and germline transmission of multiple gene mutations in rat using CRISPR-Cas systems
283 Citations Source Cite
Cited By584
Newest
Published on Jan 31, 2019in Scientific Reports 4.12
Leila Ghanbari1
Estimated H-index: 1
(University of Minnesota),
Mathew L. Rynes1
Estimated H-index: 1
(University of Minnesota)
+ 5 AuthorsSuhasa B. Kodandaramaiah8
Estimated H-index: 8
(University of Minnesota)
Over the last few decades, a plethora of tools has been developed for neuroscientists to interface with the brain. Implementing these tools requires precisely removing sections of the skull to access the brain. These delicate cranial microsurgical procedures need to be performed on the sub-millimeter thick bone without damaging the underlying tissue and therefore, require significant training. Automating some of these procedures would not only enable more precise microsurgical operations, but al...
1 Citations Source Cite
Published on Mar 5, 2019in Scientific Reports 4.12
Amélie Rezza , Christelle Jacquet1
Estimated H-index: 1
+ 8 AuthorsAlexandre Fraichard7
Estimated H-index: 7
The CRISPR/Cas9 gene editing tool enables accessible and efficient modifications which (re)ignited molecular research in certain species. However, targeted integration of large DNA fragments using CRISPR/Cas9 can still be challenging in numerous models. To systematically compare CRISPR/Cas9’s efficiency to classical homologous recombination (cHR) for insertion of large DNA fragments, we thoroughly performed and analyzed 221 experiments targeting 128 loci in mouse ES cells. Although both technolo...
Source Cite
Published on Feb 5, 2019in Cell discovery
Xiuyun Sun4
Estimated H-index: 4
(Tsinghua University),
Jun Wang7
Estimated H-index: 7
(Chinese Academy of Sciences)
+ 12 AuthorsQiuye Zhao2
Estimated H-index: 2
(Tsinghua University)
Although conventional genetic modification approaches for protein knockdown work very successfully due to the increasing use of CRISPR/Cas9, effective techniques for achieving protein depletion in adult animals, especially in large animals such as non-human primates, are lacking. Here, we report a chemical approach based on PROTACs technology that efficiently and quickly knocks down FKBP12 (12-kDa FK506-binding) protein globally in vivo. Both intraperitoneal and oral administration led to rapid,...
2 Citations Source Cite
Published on Jan 15, 2019in BMC Biotechnology 2.60
Chu Chu1
Estimated H-index: 1
(Kunming University of Science and Technology),
Zhaohui Yang1
Estimated H-index: 1
(Kunming University of Science and Technology)
+ 7 AuthorsYuyu Niu15
Estimated H-index: 15
(Kunming University of Science and Technology)
Background Non-human primate (NHP) models can closely mimic human physiological functions and are therefore highly valuable in biomedical research. Genome editing is now developing rapidly due to the precision and efficiency offered by engineered site-specific endonuclease-based systems, such as transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein-9 nuclease (Cas9) system. It has been demonst...
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Published on Jul 1, 2019in Theriogenology 2.14
Ju Zhang1
Estimated H-index: 1
(Inner Mongolia University),
Jian Liu (Inner Mongolia University)+ 8 AuthorsHao Liang2
Estimated H-index: 2
(Inner Mongolia University)
Abstract The genome editors CRISPR/Cas9 (clustered regularly interspaced short palindromicrepeats/Cas9 nuclease-null) and TALENs (transcription activator-like effector nuclease) are popularly used for targeted modification of the mammalian genome. To date, few comparative studies have been carried out to investigate the differences between the use of CRISPR/Cas9 and TALENs in genome editing for goat breeding. Here, we compared CRISPR/Cas9 and TALEN technologies at multiple levels for generating ...
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Published on Jul 1, 2019in Electronic Journal of Biotechnology 1.88
Lamei Wang1
Estimated H-index: 1
,
Sen Ma2
Estimated H-index: 2
+ 2 AuthorsYulin Chen14
Estimated H-index: 14
Source Cite
Published on Jun 5, 2019in Molecular Imaging and Biology 3.63
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Published on Feb 20, 2019in Journal of Plant Biochemistry and Biotechnology 0.77
Ajay Prakash Uniyal1
Estimated H-index: 1
(Central University of Punjab),
Sudesh Kumar Yadav25
Estimated H-index: 25
,
Vinay Kumar1
Estimated H-index: 1
(Central University of Punjab)
The CRISPR–Cas9 is emerging genome editing tool and very easy and straightforward in operation that has been tested and explored for introduction of new traits in plant systems. Recently, a number of reports have documented utilization of this technology for providing tolerance against viral diseases mediated by begomoviruses. Begomoviruses infect dicot and are transmitted by white flies and cause devastating losses to yield of important agricultural crops including tomato, cassava and cotton. A...
2 Citations Source Cite
Published on Jun 1, 2019in Seminars in Cell & Developmental Biology 6.14
William Wang1
Estimated H-index: 1
(Fujian Medical University),
Linlin Zhang2
Estimated H-index: 2
(Fudan University)
+ 1 AuthorsYiming Zeng1
Estimated H-index: 1
(Fujian Medical University)
Abstract The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) system is a prokaryotic immune system that used to resist foreign genetic factors. It rapidly becomes the hot technology in life sciences and is applies for genome editing to solve the problem of genome-derived diseases. Using CRISPR/Cas technique, the biological DNA sequence can be repaired, cut, replaced, or added. It can effectively change the human stem cells and is expected to ach...
1 Citations Source Cite