Crispr Gene Editing: Methods and Protocols (Hardcover, 2019)

Part I: Methods for CRISPR-gRNA Design and Quantification of Activity 1. CRISPR-gRNA Design Maria Pallares Masmitja, Nastassia Johanna Knodlseder, 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 Konig, 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 Ryø, 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 Dagnæs-Hansen, and Morten Frodin 21. CRISPR-Based Lentiviral Knockout Libraries for Functional Genomic Screening and Identification of Phenotype-Related Genes Emil Aagaard Thomsen and Jacob Giehm Mikkelsen