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This volume explores base editors (BEs), an invaluable CRISPR-based genome editing tool with a wide variety of versatile applications. Beginning with an overview of BEs, their diverse variants, and computational tools, the book continues with experimental applications of BEs for disease modeling in mammalian cells and generating mutagenic mice, therapeutic base editing strategies, which covers delivery methods of BE-encoded DNA plasmids, mRNAs, or ribonucleoproteins through viruses or non-viral lipid nanoparticles, and lastly, the use of BEs in plants and bacteria. 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Base Editors: Methods and Protocols serves as an ideal guide for researchers looking to use base editors to continue their studies in an array of fields.
Overview of BEs, Diverse Variants of BEs, and Computational Tools for BEs
Introduction and Perspectives of DNA Base Editors
Web-Based Computational Tools for Base Editors
Prediction of Base Editing Efficiencies and Outcomes Using DeepABE and DeepCBE
Profiling Genome-Wide Specificity of dCpf1 Cytidine Base Editors Using Digenome-Seq
Application of BEs for Disease Modeling
Base Editing of Human Hematopoietic Stem Cells
A/​C Simultaneous Conversion Using the Dual Base Editor in Human Cells
Functional Analysis of Variants in BRCA1 Using CRISPR Base Editors
Use of the Representative Base Editing Tool Target-AID to Introduce Pathogenic Mutations into Mice
Targeted Mutagenesis in Mice Using a Base Editor
Therapeutic Base Editing Strategies
Heterologous Expression and Purification of a CRISPR-Cas9-Based Adenine Base Editor
Delivering Base Editors In Vivo by Adeno-Associated Virus Vectors
The Delivery of ABE mRNA to the Adult Murine Liver by Lipid Nanoparticles (LNPs)
Ex Vivo Base Editing Therapy with Chemically Derived Hepatic Progenitors
Application of Base Editor-Mediated Genome Editing in Mouse Retina
Use of BEs in Plants and Bacteria
High-Throughput Base Editing-Mediated Artificial Evolution Streamlines Trait Gene Identification in Rice
Heritable Virus-Induced Genome Editing (VIGE) in Nicotiana attenuata
Cytosine Base Editing in Bacteria