We are analyzing https://link.springer.com/protocol/10.1007/978-1-4939-1862-1_10.

Title:
Genome Engineering Using CRISPR-Cas9 System | SpringerLink
Description:
The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 system is an adaptive immune system that exists in a variety of microbes. It could be engineered to function in eukaryotic cells as a fast, low-cost, efficient, and scalable tool for...
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Keywords {🔍}

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Topics {✒️}

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Schema {🗺️}

ScholarlyArticle:
      headline:Genome Engineering Using CRISPR-Cas9 System
      pageEnd:217
      pageStart:197
      image:https://media.springernature.com/w153/springer-static/cover/book/978-1-4939-1862-1.jpg
      genre:
         Springer Protocols
      isPartOf:
         name:Chromosomal Mutagenesis
         isbn:
            978-1-4939-1862-1
            978-1-4939-1861-4
         type:Book
      publisher:
         name:Springer New York
         logo:
            url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
            type:ImageObject
         type:Organization
      author:
            name:Le Cong
            affiliation:
                  name:Broad Institute of MIT and Harvard
                  address:
                     name:Broad Institute of MIT and Harvard, Cambridge, USA
                     type:PostalAddress
                  type:Organization
                  name:Department of Biological Engineering, Massachusetts Institute of Technology
                  address:
                     name:McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA
                     type:PostalAddress
                  type:Organization
            email:[email protected]
            type:Person
            name:Feng Zhang
            affiliation:
                  name:Broad Institute of MIT and Harvard
                  address:
                     name:Broad Institute of MIT and Harvard, Cambridge, USA
                     type:PostalAddress
                  type:Organization
                  name:Department of Biological Engineering, Massachusetts Institute of Technology
                  address:
                     name:McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA
                     type:PostalAddress
                  type:Organization
            type:Person
      keywords:Genome engineering, Cas9, CRISPR, CRISPR-Cas9, PAM, Guide RNA, sgRNA, DNA cleavage, Mutagenesis, Homologous recombination, Streptococcus pyogenes
      description:The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 system is an adaptive immune system that exists in a variety of microbes. It could be engineered to function in eukaryotic cells as a fast, low-cost, efficient, and scalable tool for manipulating genomic sequences. In this chapter, detailed protocols are described for harnessing the CRISPR-Cas9 system from Streptococcus pyogenes to enable RNA-guided genome engineering applications in mammalian cells. We present all relevant methods including the initial site selection, molecular cloning, delivery of guide RNAs (gRNAs) and Cas9 into mammalian cells, verification of target cleavage, and assays for detecting genomic modification including indels and homologous recombination. These tools provide researchers with new instruments that accelerate both forward and reverse genetics efforts.
      datePublished:2015
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         978-1-4939-1861-4
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         type:ImageObject
      name:Broad Institute of MIT and Harvard
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      name:Department of Biological Engineering, Massachusetts Institute of Technology
      address:
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         type:PostalAddress
      name:Broad Institute of MIT and Harvard
      address:
         name:Broad Institute of MIT and Harvard, Cambridge, USA
         type:PostalAddress
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      affiliation:
            name:Broad Institute of MIT and Harvard
            address:
               name:Broad Institute of MIT and Harvard, Cambridge, USA
               type:PostalAddress
            type:Organization
            name:Department of Biological Engineering, Massachusetts Institute of Technology
            address:
               name:McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA
               type:PostalAddress
            type:Organization
      email:[email protected]
      name:Feng Zhang
      affiliation:
            name:Broad Institute of MIT and Harvard
            address:
               name:Broad Institute of MIT and Harvard, Cambridge, USA
               type:PostalAddress
            type:Organization
            name:Department of Biological Engineering, Massachusetts Institute of Technology
            address:
               name:McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA
               type:PostalAddress
            type:Organization
PostalAddress:
      name:Broad Institute of MIT and Harvard, Cambridge, USA
      name:McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA
      name:Broad Institute of MIT and Harvard, Cambridge, USA
      name:McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA
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