Part I: Methods for CRISPR-gRNA Design and Quantification of Activity
1. CRISPR-gRNA Design
Maria Pallares Masmitja, Nastassia Johanna Knoedlseder, and Marc Guell
2. Tracking CRISPR's Footprints
Lin Lin and Yonglun Luo
3. Rapid Quantitative Evaluation of CRISPR Genome Editing by TIDE and TIDER
Eva Karina Brinkman and Bas van Steensel
4. Fast and Quantitative Identification of Ex Vivo Precise Genome Targeting-Induced Indel Events by IDAA
Saskia Koenig, Yang Zhang, Hans Heugh Wandall, Claudio Mussolino, and Eric Paul Bennett
5. Functional Evaluation of CRISPR Activity by the Dual-Fluorescent Surrogate System: C-Check
Lin Lin and Yonglun Luo
Part II: Methods for CRISPR Delivery
6. CRISPR-Cas9 Delivery by Artificial Virus (RRPHC)
Suleixin Yang, Qinjie Wu, Yuquan Wei, and Changyang Gong
7. Production and Validation of Lentiviral Vectors for CRISPR/Cas9 Delivery
Laura Barrett Ryo, Emil Aagaard Thomsen, and Jacob Giehm Mikkelsen
8. Rapid and Simple Screening of CRISPR Guide RNAs (gRNAs) in Cultured Cells Using Adeno-Associated Viral (AAV) Vectors
Julia Fakhiri, Manuela Nickl, and Dirk Grimm
9. Electroporation-Based CRISPR/Cas9 Gene Editing Using Cas9 Protein and Chemically Modified sgRNAs
Anders Laustsen and Rasmus O. Bak
Part III: CRISPR Gene Editing in Human iPSCs
10. Efficient Gene Editing of Human Induced Pluripotent Stem Cells Using CRISPR/Cas9
Saniye Yumlu, Sanum Bashir, Jurgen Stumm, and Ralf Kuhn
11. Editing the Genome of Human Induced Pluripotent Stem Cells Using CRISPR/Cas9 Ribonucleoprotein Complexes
Michaela Bruntraeger, Meg Byrne, Kathleen Long, and Andrew R. Bassett
12. Conditional Gene Knockout in Human Cells with Inducible CRISPR/Cas9
Kirsten E. Snijders, James D. Cooper, Ludovic Vallier, and Alessandro Bertero
Part IV: CRISPR Gene Editing in Other Cell Types
13. CRISPR/Cas9 as a Genome Editing Tool for Targeted Gene Integration in CHO Cells
Daria Sergeeva, Jose Manuel Camacho-Zaragoza, Jae Seong Lee, and Helene Faustrup Kildegaard
14. Rapid and Efficient Gene Deletion by CRISPR/Cas9
Signe Neldeborg, Lin Lin, Magnus Stougaard, and Yonglun Luo
15. Genome Editing in Mice
Lisbeth Ahm Hansen and Ernst-Martin Fuchtbauer
16. CRISPR/Cas9-Mediated Gene Tagging: A Step-by-Step Protocol
Xi Xiang, Conghui Li, Xi Chen, Hongwei Dou, Yong Li, Xiuqing Zhang, and Yonglun Luo
17. Gene Editing in Primary Cells of Cattle and Pig
Petra Vochozkova, Kilian Simmet, Eva-Maria Jemiller, Annegret Wunsch, and Nikolai Klymiuk
Part V: CRISPR Gene Therapy and Screening
18. Toward In Vivo Gene Therapy Using CRISPR
Kristian Alsbjerg Skipper and Jacob Giehm Mikkelsen
19. CRISPR Gene Therapy of the Eye: Targeted Knockout of Vegfa in Mouse Retina by Lentiviral Delivery
Andreas Holmgaard, Sidsel Alsing, Anne Louise Askou, and Thomas J. Corydon
20. In Vivo Editing of the Adult Mouse Liver Using CRISPR/Cas9 and Hydrodynamic Tail Vein Injection
Francesco Niola, Frederik Dagnaes-Hansen, and Morten Froedin
21. CRISPR-Based Lentiviral Knockout Libraries for Functional Genomic Screening and Identification of Phenotype-Related Genes
Emil Aagaard Thomsen and Jacob Giehm Mikkelsen
