Crispr Gene Editing: Methods and Protocols

ISBN-13: 9781493991693 / Angielski / Twarda / 2019 / 362 str.

Yonglun Luo

Spis treści

Part I: Methods for CRISPR-gRNA Design and Quantification of Activity

 

1. CRISPR-gRNA Design

            Maria Pallarès Masmitjà, Nastassia Johanna Knödlseder, and Marc Güell

 

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 König, 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, Jürgen Stumm, and Ralf Kühn

 

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 Füchtbauer

 

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 Wünsch, 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 Frödin

 

21. CRISPR-Based Lentiviral Knockout Libraries for Functional Genomic Screening and Identification of Phenotype-Related Genes

            Emil Aagaard Thomsen and Jacob Giehm Mikkelsen

Blurb

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.