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CRISPR Gene Editing

Methods and Protocols

  • Book
  • © 2019

Overview

Editors:
  1. Yonglun Luo

    1. Department of Biomedicine, Aarhus University, Aarhus, Denmark

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  • Includes cutting-edge techniques
  • Provides step-by-step detail essential for reproducible results
  • Contains key implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 1961)

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Table of contents (21 protocols)

  1. Front Matter

    Pages i-xi
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  2. Methods for CRISPR-gRNA Design and Quantification of Activity

    1. Front Matter

      Pages 1-1
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    2. CRISPR-gRNA Design

      • Maria Pallarès Masmitjà, Nastassia Knödlseder, Marc Güell
      Pages 3-11

    3. Tracking CRISPR’s Footprints

      • Lin Lin, Yonglun Luo
      Pages 13-28

    4. Rapid Quantitative Evaluation of CRISPR Genome Editing by TIDE and TIDER

      • Eva Karina Brinkman, Bas van Steensel
      Pages 29-44Open Access

    5. Fast and Quantitative Identification of Ex Vivo Precise Genome Targeting-Induced Indel Events by IDAA

      • Saskia König, Zhang Yang, Hans Heugh Wandall, Claudio Mussolino, Eric Paul Bennett
      Pages 45-66

    6. Functional Evaluation of CRISPR Activity by the Dual-Fluorescent Surrogate System: C-Check

      • Lin Lin, Yonglun Luo
      Pages 67-77

  3. Methods for CRISPR Delivery

    1. Front Matter

      Pages 79-79
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    2. CRISPR-Cas9 Delivery by Artificial Virus (RRPHC)

      • Suleixin Yang, Qinjie Wu, Yuquan Wei, Changyang Gong
      Pages 81-91

    3. Production and Validation of Lentiviral Vectors for CRISPR/Cas9 Delivery

      • Laura Barrett Ryø, Emil Aagaard Thomsen, Jacob Giehm Mikkelsen
      Pages 93-109

    4. Rapid and Simple Screening of CRISPR Guide RNAs (gRNAs) in Cultured Cells Using Adeno-Associated Viral (AAV) Vectors

      • Julia Fakhiri, Manuela Nickl, Dirk Grimm
      Pages 111-126

    5. Electroporation-Based CRISPR/Cas9 Gene Editing Using Cas9 Protein and Chemically Modified sgRNAs

      • Anders Laustsen, Rasmus O. Bak
      Pages 127-134

  4. CRISPR Gene Editing in Human iPSCs

    1. Front Matter

      Pages 135-135
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    2. Efficient Gene Editing of Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

      • Saniye Yumlu, Sanum Bashir, Jürgen Stumm, Ralf Kühn
      Pages 137-151

    3. Editing the Genome of Human Induced Pluripotent Stem Cells Using CRISPR/Cas9 Ribonucleoprotein Complexes

      • Michaela Bruntraeger, Meg Byrne, Kathleen Long, Andrew R. Bassett
      Pages 153-183

    4. Conditional Gene Knockout in Human Cells with Inducible CRISPR/Cas9

      • Kirsten E. Snijders, James D. Cooper, Ludovic Vallier, Alessandro Bertero
      Pages 185-209

  5. CRISPR Gene Editing in Other Cell Types

    1. Front Matter

      Pages 211-211
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    2. CRISPR/Cas9 as a Genome Editing Tool for Targeted Gene Integration in CHO Cells

      • Daria Sergeeva, Jose Manuel Camacho-Zaragoza, Jae Seong Lee, Helene Faustrup Kildegaard
      Pages 213-232

    3. Rapid and Efficient Gene Deletion by CRISPR/Cas9

      • Signe Neldeborg, Lin Lin, Magnus Stougaard, Yonglun Luo
      Pages 233-247

    4. Genome Editing in Mice

      • Lisbeth Ahm Hansen, Ernst-Martin Füchtbauer
      Pages 249-254
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Keywords

About this book

This detailed volume guides readers through strategic planning and user-friendly guidelines in order to select the most suitable CRISPR-Cas system and target sites with high activity and specificity. Methods covering CRISPR gRNA design, CRISPR delivery, CRISPR activity quantification (indel quantification), and examples of applying CRISPR gene editing in human pluripotent stem cells, primary cells, gene therapy, and genetic screening are included. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and invaluable, CRISPR Gene Editing: Methods and Protocols will assist undergraduates, graduates, and researchers with detailed guidelines and methods for the vitally important CRISPR gene editing field. 


Chapter 3 is available open access under a CC BY 4.0 license via link.springer.com.

Editors and Affiliations

  • Department of Biomedicine, Aarhus University, Aarhus, Denmark

    Yonglun Luo

Bibliographic Information

  • Book Title: CRISPR Gene Editing

  • Book Subtitle: Methods and Protocols

  • Editors: Yonglun Luo

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-9170-9

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2019

  • Hardcover ISBN: 978-1-4939-9169-3Published: 26 March 2019

  • eBook ISBN: 978-1-4939-9170-9Published: 25 March 2019

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 362

  • Number of Illustrations: 15 b/w illustrations, 61 illustrations in colour

  • Topics: Human Genetics, Genetic Engineering

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