ENGLISH

CRISPR Gene Editing: Methods and Protocols

By Yonglun Luo

Book information

Publisher
Springer New York; Humana
Year
2019
ISBN
978-1-4939-9169-3;978-1-4939-9170-9
Language
english
Format
PDF
Filesize
10 MB (10343837 bytes)
Series
Methods in Molecular Biology 1961
Edition
1st ed.
Pages
XI, 362\356
Time added
2019-09-18 12:16:59
Biomedicine; Human Genetics; Genetic Engineering

Description

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. Front Matter ....Pages i-xi Front Matter ....Pages 1-1 CRISPR-gRNA Design (Maria Pallarès Masmitjà, Nastassia Knödlseder, Marc Güell)....Pages 3-11 Tracking CRISPR’s Footprints (Lin Lin, Yonglun Luo)....Pages 13-28 Rapid Quantitative Evaluation of CRISPR Genome Editing by TIDE and TIDER (Eva Karina Brinkman, Bas van Steensel)....Pages 29-44 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 Functional Evaluation of CRISPR Activity by the Dual-Fluorescent Surrogate System: C-Check (Lin Lin, Yonglun Luo)....Pages 67-77 Front Matter ....Pages 79-79 CRISPR-Cas9 Delivery by Artificial Virus (RRPHC) (Suleixin Yang, Qinjie Wu, Yuquan Wei, Changyang Gong)....Pages 81-91 Production and Validation of Lentiviral Vectors for CRISPR/Cas9 Delivery (Laura Barrett Ryø, Emil Aagaard Thomsen, Jacob Giehm Mikkelsen)....Pages 93-109 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 Electroporation-Based CRISPR/Cas9 Gene Editing Using Cas9 Protein and Chemically Modified sgRNAs (Anders Laustsen, Rasmus O. Bak)....Pages 127-134 Front Matter ....Pages 135-135 Efficient Gene Editing of Human Induced Pluripotent Stem Cells Using CRISPR/Cas9 (Saniye Yumlu, Sanum Bashir, Jürgen Stumm, Ralf Kühn)....Pages 137-151 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 Conditional Gene Knockout in Human Cells with Inducible CRISPR/Cas9 (Kirsten E. Snijders, James D. Cooper, Ludovic Vallier, Alessandro Bertero)....Pages 185-209 Front Matter ....Pages 211-211 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 Rapid and Efficient Gene Deletion by CRISPR/Cas9 (Signe Neldeborg, Lin Lin, Magnus Stougaard, Yonglun Luo)....Pages 233-247 Genome Editing in Mice (Lisbeth Ahm Hansen, Ernst-Martin Füchtbauer)....Pages 249-254 CRISPR/Cas9-Mediated Gene Tagging: A Step-by-Step Protocol (Xi Xiang, Conghui Li, Xi Chen, Hongwei Dou, Yong Li, Xiuqing Zhang et al.)....Pages 255-269 Gene Editing in Primary Cells of Cattle and Pig (Petra Vochozkova, Kilian Simmet, Eva-Maria Jemiller, Annegret Wünsch, Nikolai Klymiuk)....Pages 271-289 Front Matter ....Pages 291-291 Toward In Vivo Gene Therapy Using CRISPR (Kristian Alsbjerg Skipper, Jacob Giehm Mikkelsen)....Pages 293-306 CRISPR Gene Therapy of the Eye: Targeted Knockout of Vegfa in Mouse Retina by Lentiviral Delivery (Andreas Holmgaard, Sidsel Alsing, Anne Louise Askou, Thomas J. Corydon)....Pages 307-328 In Vivo Editing of the Adult Mouse Liver Using CRISPR/Cas9 and Hydrodynamic Tail Vein Injection (Francesco Niola, Frederik Dagnæs-Hansen, Morten Frödin)....Pages 329-341 CRISPR-Based Lentiviral Knockout Libraries for Functional Genomic Screening and Identification of Phenotype-Related Genes (Emil Aagaard Thomsen, Jacob Giehm Mikkelsen)....Pages 343-357 Back Matter ....Pages 359-362

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