Generating rats with conditional alleles using CRISPR/Cas9

Ma, Yuanwu; Zhang, Xu; Shen, Bin; Lu, Yingdong; Chen, Wei; Ma, Jing; Bai, Lin; Huang, Xingxu; Zhang, Lianfeng

doi:10.1038/cr.2013.157

Cited by 172 publications

(174 citation statements)

References 10 publications

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“…Unlike the predecessor zinc finger nuclease (ZFN) and TAL effector nuclease (TALEN), which involve dimerizing fusion proteins including the DNA binding domains of ZF and TAL and cleavage domains of FokI endonuclease, Cas9/gRNA is a ribonucleoprotein active on target DNA (Char et al 2017). This technology has been widely adopted to study important genes in the cell of mice (Mashiko et al 2014), monkeys (Niu et al 2014), and other organisms, including bacteria (Fabre et al 2014), yeast , zebrafish (Hwang et al 2013), Drosophila (Gratz et al 2014), rabbits , pigs (Hai et al 2014), rats (Ma et al 2014), human , and plants . In biomedical field, significant applications have already been achieved including correcting human genetic disorder, treatment of the acquired immune deficiency syndrome (AIDS) or promoting anti-tumor immunotherapy, and genetic manipulation of domesticated animals for production of biologic medical materials among others (Cai et al 2016).…”

Section: Introductionmentioning

confidence: 99%

CRISPR/CAS9 as a Powerful Tool for Crop Improvement

et al. 2017

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This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Recently, a modified version of CRISPR/Cas9 with added novel functions has been developed that enables programmable direct irreversible conversion of a target DNA base. In this review, we summarized the milestone applications of CRISPR/ Cas9 in plants with a focus on major crops. We also present the implications of an improved version of this technology in the current plant breeding programs.

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Section: Introductionmentioning

confidence: 99%

CRISPR/CAS9 as a Powerful Tool for Crop Improvement

et al. 2017

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“…The Cas9/sgRNA complex recognizes the complementary 20-nucleotide genomic sequence preceding a 5'-NGG-3' Protospacer adjacent motif (PAM) sequence in mammalian cells 22,23 . It has been successfully used for effective generation of genetically modified cells, viruses and animal models [22][23][24][25][26][27][28][29][30][31][32] .…”

Section: Introductionmentioning

confidence: 99%

A Simple and Efficient Approach to Construct Mutant Vaccinia Virus Vectors

Yuan

Wang

Chard

et al. 2016

JoVE

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The CRISPR-associated endonuclease Cas9 can edit particular genomic loci directed by a single guide RNA (gRNA). The CRISPR/Cas9 system has been successfully employed for editing genomes of various organisms. Here we describe a protocol for editing the vaccinia virus (VV) genome in the cytoplasm of VV-infected CV-1 cells using the RNA-guided Cas9. RNA-guided Cas9 induces double-stranded DNA breaks facilitating homologous recombination efficiently and specifically in the targeted site of VV and a transgene can be incorporated into these sites by homologous recombination. By using a site-specific homologous vector with transgene(s), a N1L gene-deleted VV with the red fluorescence protein (RFP) gene incorporated in this region was generated with a successful recombination efficiency 10 times greater than that obtained from the conventional homologous recombination method. This protocol demonstrates successful use of RNA-guided Cas9 system to generate mutant VVs with enhanced efficiency.

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“…For instance, its efficiency is higher than HR, which makes it feasible to modify human cells; and it is also considerably easier to design guide RNA constructs than ZFNs and TALENs, making it suitable to perform genome-scale genes knockout screening. The gene knockout capabilities of the CRISPR-Cas9 system have been successfully shown in many model organisms, such as mice Wu et al 2013;Yang et al 2013b), rats (Hu et al 2013;Li et al 2013;Ma et al 2014), pigs (Hai et al 2014), and even monkeys . The CRISPR-Cas9 system can be used to correct a genetic disease in mouse via HDR based on an exogenously supplied oligonucleotide or the endogenous Wilms' tumor (WT) allele by coinjection into zygotes of Cas9 mRNA and an sgRNA targeting the mutant allele .…”

Section: Genome-wide Screening Via Crispr-cas9 For Diploid Pluripotenmentioning

confidence: 99%

Next-Generation Models of Human Cardiogenesis via Genome Editing

Lian

et al. 2014

Cold Spring Harbor Perspectives in Medicine

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Cardiogenesis is one of the earliest and most important steps during human development and is orchestrated by discrete families of heart progenitors, which build distinct regions of the fetal heart. For the past decade, a lineage map for the distinct subsets of progenitors that generate the embryonic mammalian heart has begun to lay a foundation for the development of new strategies for rebuilding the adult heart after injury, an unmet clinical need for the vast majority of patients with end-stage heart failure who are not heart transplant recipients. The studies also have implications for the root causes of congenital heart disease, which affects 1 in 50 live births, the most prevalent malformations in children. Although much of this insight has been generated in murine models, it is becoming increasingly clear that there can be important divergence with principles and pathways for human cardiogenesis, as well as for regenerative pathways. The development of human stem cell models, coupled with recent advances in genome editing with RNA-guided endonucleases, offers a new approach for the primary study of human cardiogenesis. In addition, application of the technology to the in vivo setting in large animal models, including nonhuman primates, has opened the door to genome-edited large animal models of adult and congenital heart disease, as well as a detailed mechanistic dissection of the more diverse and complex set of progenitor families and pathways, which guide human cardiogenesis. Implications of this new technology for a new generation of human-based, genetically tractable systems are discussed, along with potential therapeutic applications. A DECADE OF ADVANCES IN MURINE CARDIOGENESISO ver the past decade, major advances in our understanding of conserved principles and pathways for cardiogenesis have been made through genetically engineered mouse models of cardiogenesis. The assumption has been that the major underpinnings of such a conserved and essential organ system would have parallels in human organogenesis. The approach has been largely based on gene knockout apEditors:

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Generating rats with conditional alleles using CRISPR/Cas9

Cited by 172 publications

References 10 publications

CRISPR/CAS9 as a Powerful Tool for Crop Improvement

CRISPR/CAS9 as a Powerful Tool for Crop Improvement

A Simple and Efficient Approach to Construct Mutant Vaccinia Virus Vectors

Next-Generation Models of Human Cardiogenesis via Genome Editing

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