CRISPR Gene Editing: Methods and Protocols

✍ Scribed by Yonglun Luo

Publisher: Springer New York; Humana
Year: 2019
Tongue: English
Leaves: 356
Series: Methods in Molecular Biology 1961
Edition: 1st ed.
Category: Library

No coin nor oath required. For personal study only.

✦ Synopsis

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.

✦ Table of Contents

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

✦ Subjects

Biomedicine; Human Genetics; Genetic Engineering

📜 SIMILAR VOLUMES

CRISPR: Methods and Protocols

📁 CRISPR: Methods and Protocols

✍ Magnus Lundgren, Emmanuelle Charpentier, Peter C. Fineran 📂 Library 📅 2015 🏛 Humana Press 🌐 English

This volume presents a list of cutting-edge protocols for the study of CRISPR-Cas defense systems and their applications at the genomic, genetic, biochemical and structural levels. CRISPR: Methods and Protocols guides readers through techniques that have been developed specifically for the

CRISPR: Methods and Protocols

📁 CRISPR: Methods and Protocols

✍ Magnus Lundgren, Emmanuelle Charpentier, Peter C. Fineran (eds.) 📂 Library 📅 2015 🏛 Humana Press 🌐 English

This volume presents a list of cutting-edge protocols for the study of CRISPR-Cas defense systems and their applications at the genomic, genetic, biochemical and structural levels. CRISPR: Methods and Protocols guides readers through techniques that have been developed specifically for

Plant Genome Editing with CRISPR Systems

📁 Plant Genome Editing with CRISPR Systems: Methods and Protocols

✍ Yiping Qi 📂 Library 📅 2019 🏛 Springer New York;Humana Press 🌐 English

This volume provides readers with wide-ranging coverage of CRISPR systems and their applications in various plant species. The chapters in this book discuss topics such as plant DNA repair and genome editing; analysis of CRISPR-induced mutations; multiplexed CRISPR/Cas9 systems; CRISPR-Cas12a

CRISPR Guide RNA Design: Methods and Pro

📁 CRISPR Guide RNA Design: Methods and Protocols

✍ Tudor A. Fulga, David J. H. F. Knapp, Quentin R. V. Ferry 📂 Library 📅 2021 🏛 Springer US;Humana 🌐 English

This detailed volume focuses on the CRISPR-associated guide RNA and how it can be designed, modified, and validated for a broad repertoire of purposes. Beginning with a section on computational design of target-specific guide RNAs, the book continues by covering chemical modifications to alter gu

Rice Genome Engineering and Gene Editing

📁 Rice Genome Engineering and Gene Editing: Methods and Protocols (Methods in Molecular Biology, 2238)

✍ Anindya Bandyopadhyay (editor), Roger Thilmony (editor) 📂 Library 📅 2021 🏛 Humana 🌐 English

This detailed volume explores rice molecular biology, genetic engineering, and genome editing technologies. Dividing into three parts, the book covers subjects such as genetic engineering and tissue culture of rice, including efficient methods for rice transformation and regeneration, genome e

Gene Synthesis: Methods and Protocols

📁 Gene Synthesis: Methods and Protocols

✍ Julie A. Marchand, Jean Peccoud (auth.), Jean Peccoud (eds.) 📂 Library 📅 2012 🏛 Humana Press 🌐 English

The de novo fabrication of custom DNA molecules is a transformative technology that significantly affects the biotechnology industry. Basic genetic engineering techniques for manipulating DNA in vitro opened an incredible field of opportunity in the life sciences. In, Gene Syn