The Experts below are selected from a list of 16965 Experts worldwide ranked by ideXlab platform

Caixia Gao - One of the best experts on this subject based on the ideXlab platform.

  • genome editing in rice and wheat using the CRISPR Cas System
    Nature Protocols, 2014
    Co-Authors: Qiwei Shan, Yanpeng Wang, Caixia Gao
    Abstract:

    The Gao laboratory provides its protocol for targeted editing of crop genomes using the CRISPR/Cas System. Genome alterations can be induced by transient transfection of rice or wheat protoplasts or by stable transformation of rice plants.

  • Targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas System
    Journal of Genetics and Genomics, 2014
    Co-Authors: Zhen Liang, Kang Zhang, Kunling Chen, Caixia Gao
    Abstract:

    Transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) Systems have emerged as powerful tools for genome editing in a variety of species. Here, we report, for the first time, targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas System. We designed five TALENs targeting 4 genes, namely ZmPDS, ZmIPK1A, ZmIPK, ZmMRP4, and obtained targeting efficiencies of up to 23.1% in protoplasts, and about 13.3% to 39.1% of the transgenic plants were somatic mutations. Also, we constructed two gRNAs targeting the ZmIPK gene in maize protoplasts, at frequencies of 16.4% and 19.1%, respectively. In addition, the CRISPR/Cas System induced targeted mutations in Z. mays protoplasts with efficiencies (13.1%) similar to those obtained with TALENs (9.1%). Our results show that both TALENs and the CRISPR/Cas System can be used for genome modification in maize. © 2013.

Huimin Zhao - One of the best experts on this subject based on the ideXlab platform.

  • high efficiency genome editing of streptomyces species by an engineered CRISPR Cas System
    Methods in Enzymology, 2016
    Co-Authors: Yuhang Wang, Ryan E Cobb, Huimin Zhao
    Abstract:

    Next-generation sequencing technologies have rapidly expanded the genomic information of numerous organisms and revealed a rich reservoir of natural product gene clusters from microbial genomes, especially from Streptomyces, the largest genus of known actinobacteria at present. However, genetic engineering of these bacteria is often time consuming and labor intensive, if even possible. In this chapter, we describe the design and construction of pCRISPomyces, an engineered Type II CRISPR/Cas System, for targeted multiplex gene deletions in Streptomyces lividans, Streptomyces albus, and Streptomyces viridochromogenes with editing efficiency ranging from 70% to 100%. We demonstrate pCRISPomyces as a powerful tool for genome editing in Streptomyces.

  • high efficiency multiplex genome editing of streptomyces species using an engineered CRISPR Cas System
    ACS Synthetic Biology, 2015
    Co-Authors: Ryan E Cobb, Yajie Wang, Huimin Zhao
    Abstract:

    Actinobacteria, particularly those of genus Streptomyces, remain invaluable hosts for the discovery and engineering of natural products and their cognate biosynthetic pathways. However, genetic manipulation of these bacteria is often labor and time intensive. Here, we present an engineered CRISPR/Cas System for rapid multiplex genome editing of Streptomyces strains, demonstrating targeted chromosomal deletions in three different Streptomyces species and of various sizes (ranging from 20 bp to 30 kb) with efficiency ranging from 70 to 100%. The designed pCRISPomyces plasmids are amenable to assembly of spacers and editing templates via Golden Gate assembly and isothermal assembly (or traditional digestion/ligation), respectively, allowing rapid plasmid construction to target any genomic locus of interest. As such, the pCRISPomyces System represents a powerful new tool for genome editing in Streptomyces.

Kunling Chen - One of the best experts on this subject based on the ideXlab platform.

  • targeted mutagenesis in zea mays using talens and the CRISPR Cas System
    Journal of Genetics and Genomics, 2014
    Co-Authors: Zhen Liang, Kang Zhang, Kunling Chen
    Abstract:

    Abstract Transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) Systems have emerged as powerful tools for genome editing in a variety of species. Here, we report, for the first time, targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas System. We designed five TALENs targeting 4 genes, namely ZmPDS, ZmIPK1A, ZmIPK, ZmMRP4, and obtained targeting efficiencies of up to 23.1% in protoplasts, and about 13.3% to 39.1% of the transgenic plants were somatic mutations. Also, we constructed two gRNAs targeting the ZmIPK gene in maize protoplasts, at frequencies of 16.4% and 19.1%, respectively. In addition, the CRISPR/Cas System induced targeted mutations in Z. mays protoplasts with efficiencies (13.1%) similar to those obtained with TALENs (9.1%). Our results show that both TALENs and the CRISPR/Cas System can be used for genome modification in maize.

  • Targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas System
    Journal of Genetics and Genomics, 2014
    Co-Authors: Zhen Liang, Kang Zhang, Kunling Chen, Caixia Gao
    Abstract:

    Transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) Systems have emerged as powerful tools for genome editing in a variety of species. Here, we report, for the first time, targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas System. We designed five TALENs targeting 4 genes, namely ZmPDS, ZmIPK1A, ZmIPK, ZmMRP4, and obtained targeting efficiencies of up to 23.1% in protoplasts, and about 13.3% to 39.1% of the transgenic plants were somatic mutations. Also, we constructed two gRNAs targeting the ZmIPK gene in maize protoplasts, at frequencies of 16.4% and 19.1%, respectively. In addition, the CRISPR/Cas System induced targeted mutations in Z. mays protoplasts with efficiencies (13.1%) similar to those obtained with TALENs (9.1%). Our results show that both TALENs and the CRISPR/Cas System can be used for genome modification in maize. © 2013.

Lluis Montoliu - One of the best experts on this subject based on the ideXlab platform.

  • The new CRISPR–Cas System: RNA-guided genome engineering to efficiently produce any desired genetic alteration in animals
    Transgenic Research, 2014
    Co-Authors: Davide Seruggia, Lluis Montoliu
    Abstract:

    The CRISPR–Cas System is the newest targeted nuclease for genome engineering. In less than 1 year, the ease, robustness and efficiency of this method have facilitated an immense range of genetic modifications in most model organisms. Full and conditional gene knock-outs, knock-ins, large chromosomal deletions and subtle mutations can be obtained using combinations of clustered regularly interspaced short palindromic repeats (CRISPRs) and DNA donors. In addition, with CRISPR–Cas compounds, multiple genetic modifications can be introduced seamlessly in a single step. CRISPR–Cas not only brings genome engineering capacities to species such as rodents and livestock in which the existing toolbox was already large, but has also enabled precise genetic engineering of organisms with difficult-to-edit genomes such as zebrafish, and of technically challenging species such as non-human primates. The CRISPR–Cas System allows generation of targeted mutations in mice, even in laboratories with limited or no access to the complex, time-consuming standard technology using mouse embryonic stem cells. Here we summarize the distinct applications of CRISPR–Cas technology for obtaining a variety of genetic modifications in different model organisms, underlining their advantages and limitations relative to other genome editing nucleases. We will guide the reader through the many publications that have seen the light in the first year of CRISPR–Cas technology.

  • mouse genome editing using the CRISPR Cas System
    Current protocols in human genetics, 2014
    Co-Authors: Donald W Harms, Davide Seruggia, Lluis Montoliu, Rolen M Quadros, Masato Ohtsuka, Gou Takahashi, Channabasavaiah B Gurumurthy
    Abstract:

    The availability of techniques to create desired genetic mutations has enabled the laboratory mouse as an extensively used model organism in biomedical research including human genetics. A new addition to this existing technical repertoire is the CRISPR/Cas System. Specifically, this System allows editing of the mouse genome much faster than the previously used techniques and more importantly multiple mutations can be created in a single experiment. Here we provide protocols for preparation of CRISPR/Cas reagents and microinjection into one cell mouse embryos to create knockout or knock-in mouse models.

  • the new CRISPR Cas System rna guided genome engineering to efficiently produce any desired genetic alteration in animals
    Transgenic Research, 2014
    Co-Authors: Davide Seruggia, Lluis Montoliu
    Abstract:

    The CRISPR–Cas System is the newest targeted nuclease for genome engineering. In less than 1 year, the ease, robustness and efficiency of this method have facilitated an immense range of genetic modifications in most model organisms. Full and conditional gene knock-outs, knock-ins, large chromosomal deletions and subtle mutations can be obtained using combinations of clustered regularly interspaced short palindromic repeats (CRISPRs) and DNA donors. In addition, with CRISPR–Cas compounds, multiple genetic modifications can be introduced seamlessly in a single step. CRISPR–Cas not only brings genome engineering capacities to species such as rodents and livestock in which the existing toolbox was already large, but has also enabled precise genetic engineering of organisms with difficult-to-edit genomes such as zebrafish, and of technically challenging species such as non-human primates. The CRISPR–Cas System allows generation of targeted mutations in mice, even in laboratories with limited or no access to the complex, time-consuming standard technology using mouse embryonic stem cells. Here we summarize the distinct applications of CRISPR–Cas technology for obtaining a variety of genetic modifications in different model organisms, underlining their advantages and limitations relative to other genome editing nucleases. We will guide the reader through the many publications that have seen the light in the first year of CRISPR–Cas technology.

Zhen Liang - One of the best experts on this subject based on the ideXlab platform.

  • targeted mutagenesis in zea mays using talens and the CRISPR Cas System
    Journal of Genetics and Genomics, 2014
    Co-Authors: Zhen Liang, Kang Zhang, Kunling Chen
    Abstract:

    Abstract Transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) Systems have emerged as powerful tools for genome editing in a variety of species. Here, we report, for the first time, targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas System. We designed five TALENs targeting 4 genes, namely ZmPDS, ZmIPK1A, ZmIPK, ZmMRP4, and obtained targeting efficiencies of up to 23.1% in protoplasts, and about 13.3% to 39.1% of the transgenic plants were somatic mutations. Also, we constructed two gRNAs targeting the ZmIPK gene in maize protoplasts, at frequencies of 16.4% and 19.1%, respectively. In addition, the CRISPR/Cas System induced targeted mutations in Z. mays protoplasts with efficiencies (13.1%) similar to those obtained with TALENs (9.1%). Our results show that both TALENs and the CRISPR/Cas System can be used for genome modification in maize.

  • Targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas System
    Journal of Genetics and Genomics, 2014
    Co-Authors: Zhen Liang, Kang Zhang, Kunling Chen, Caixia Gao
    Abstract:

    Transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) Systems have emerged as powerful tools for genome editing in a variety of species. Here, we report, for the first time, targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas System. We designed five TALENs targeting 4 genes, namely ZmPDS, ZmIPK1A, ZmIPK, ZmMRP4, and obtained targeting efficiencies of up to 23.1% in protoplasts, and about 13.3% to 39.1% of the transgenic plants were somatic mutations. Also, we constructed two gRNAs targeting the ZmIPK gene in maize protoplasts, at frequencies of 16.4% and 19.1%, respectively. In addition, the CRISPR/Cas System induced targeted mutations in Z. mays protoplasts with efficiencies (13.1%) similar to those obtained with TALENs (9.1%). Our results show that both TALENs and the CRISPR/Cas System can be used for genome modification in maize. © 2013.