CRISPR Gene Editing

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

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.

• CRISPR-Cas
• Genome editing
• Genetic modification
• gRNA design
• Regenerative medicine
• Genetic manipulation tools
• CRISPR delivery
• Gene therapy
• genetic engineering

• Department of Biomedicine, Aarhus University, Aarhus, Denmark

  Yonglun Luo

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