Genome Editing
A Practical Guide to Research and Clinical Applications
- 1st Edition - March 5, 2021
- Imprint: Academic Press
- Author: Kiran Musunuru
- Language: English
- Paperback ISBN:9 7 8 - 0 - 1 2 - 8 2 3 4 8 4 - 6
- eBook ISBN:9 7 8 - 0 - 1 2 - 8 2 3 6 4 7 - 5
Genome Editing: A Practical Guide to Research and Clinical Applications is geared towards investigators interested in learning how to use CRISPR-Cas9-based technologies, with a… Read more
Purchase options
Institutional subscription on ScienceDirect
Request a sales quoteGenome Editing: A Practical Guide to Research and Clinical Applications is geared towards investigators interested in learning how to use CRISPR-Cas9-based technologies, with a focus on cardiovascular research and clinical applications. Covering a range of topics from the basics of genome editing to design considerations, to assessments and applications, this reference allows readers to get started and establish a full workflow from the beginning of the project to its full completion. With worked examples drawn from real-life experiments, as well as troubleshooting and pitfalls to avoid, the book serves as an essential reference for researchers and investigators in both cardiovascular and biomedical research.
- Help readers familiarise with the variety of genome-editing approaches that are being applied in cardiovascular research and medicine, i.e., both research applications and clinical applications
- Understand the use of genome editing through worked examples (based on real-life experiments) in which CRISPR-Cas9 is employed, online tools to design CRISPR-Cas9 reagents, methods to interpret data from genome-editing experiments, the downsides of genome-editing technology - both the scientific and ethical pitfalls to avoid
- Written in an easy-to-follow manner, guiding readers from the design of the project to its completion
- Includes unpublished and new methods
Investigators/researchers in cardiovascular research. (from trainees to principal investigators), clinicians, cardiologists. Investigators broadly engaged in biomedical research, advanced undergraduate, graduate students, educators and course developers who seek to stimulate interest in emerging research applications, college instructors/ professors teaching seminars on emerging research approaches. Those interested in ethics and health policy as they seek to understand the approach and applications well enough to make health policy recommendations
- Cover image
- Title page
- Table of Contents
- Copyright
- Dedication
- Preface
- Chapter 1. A brief history and primer on genome editing
- Abstract
- Homologous recombination
- Zinc-finger nucleases (ZFNs)
- Transcription activator-like effector nucleases (TALENs)
- CRISPR-Cas9 and RNA-guided endonucleases
- References
- Chapter 2. Choosing a genome editing strategy and target site
- Abstract
- Introduction
- Gene knockout with single-site targeting
- Gene knockout with double-site targeting
- Gene knockout via sequence insertion, and the problem of long noncoding RNAs
- Inserting or correcting mutations
- Inserting a gene or other DNA sequence
- References
- Chapter 3. Choosing a nuclease, guide RNA, and repair template
- Abstract
- Streptococcus pyogenes CRISPR-Cas9
- Choosing Streptococcus pyogenes Cas9 guide RNAs for non-homologous end joining
- Choosing guide RNAs and synthetic DNA repair templates for homology-directed repair
- Streptococcus pyogenes Cas9 with altered protospacer-adjacent motif preferences
- Alternative Cas9 proteins
- Cas12 proteins and other potential editors
- Afternote
- References
- Chapter 4. Assessing the outcomes of genome editing
- Abstract
- Methods to assess genome editing of bulk cells
- Methods to assess genome editing of clonal cells
- Mosaicism
- Assessing for homology-directed repair and ruling out hemizygosity
- References
- Chapter 5. Assessing for off-target mutagenesis
- Abstract
- The potential hazards of off-target mutagenesis
- Defining the extent of off-target mutagenesis
- Discovery phase
- Validation phase
- Strategies to reduce off-target mutagenesis
- Practical example
- References
- Chapter 6. Base editing
- Abstract
- The BE(x) cytosine base editors
- Other cytosine base editors
- Adenine base editors
- Mitochondrial base editors
- Off-target effects of base editors
- Base editing for gene knockout
- Base editing for inserting or correcting mutations
- References
- Chapter 7. Alternative types of editing
- Abstract
- Epigenome editing
- Practical example of epigenome editing
- Genome-wide CRISPR screens
- RNA targeting and editing
- Practical example of RNA targeting
- Prime editing
- Practical example of prime editing
- References
- Chapter 8. Genome editing for cellular disease modeling
- Abstract
- Human pluripotent stem cells and the advantages of genome editing
- Study design considerations: example #1
- Study design considerations: example #2
- Study design considerations: example #3
- Summary
- References
- Chapter 9. Genome editing for functional experiments and screens
- Abstract
- Rapid generation of knockoutand knock-in mouse models: The study of KLF14
- Combining a variety of editing approaches: The study of two lipid-associated loci
- Multiplex genome editing: Generating an allelic series of cell lines to annotate patient variants of uncertain significance
- Genome-wide CRISPR screens: Improving hepatocyte differentiation and identifying genetic modifiers of doxorubicin-induced cardiotoxicity
- References
- Chapter 10. Therapeutic genome editing
- Abstract
- Ex vivo therapeutic genome editing
- In vivo therapeutic genome editing
- Practical example
- Ethical considerations
- References
- Index
- Edition: 1
- Published: March 5, 2021
- No. of pages (Paperback): 230
- No. of pages (eBook): 230
- Imprint: Academic Press
- Language: English
- Paperback ISBN: 9780128234846
- eBook ISBN: 9780128236475
KM
Kiran Musunuru
Kiran Musunuru, MD, PhD, MPH, ML received his medical degree from Weill Cornell Medical College, his PhD from The Rockefeller University, and his Master of Public Health from Johns Hopkins Bloomberg School of Public Health. He trained in Internal Medicine at Brigham and Women’s Hospital and Cardiovascular Medicine at Johns Hopkins Hospital, followed by postdoctoral work at Massachusetts General Hospital and the Broad Institute of MIT and Harvard. Dr Musunuru’s research focuses on the genetics of cardiovascular and metabolic diseases and seeks to identify naturally occurring genetic variants that predispose to or protect against disease and can be used to develop therapies to protect the entire population. His expertise includes the use of human pluripotent stem cells as a platform for disease modelling and the use of genome-editing tools such as CRISPR-Cas9 for research and therapeutic applications. In 2016, he received the Presidential Early Career Award for Scientists and Engineers from U.S. President Obama, as well as the American Heart Association’s Award of Meritorious Achievement. He became Editor-in-Chief of Circulation: Genomic and Precision Medicine (an American Heart Association journal) in 2018.
Affiliations and expertise
Associate Professor of Cardiovascular Medicine and Genetics, Perelman School of Medicine, University of Pennsylvania, USARead Genome Editing on ScienceDirect